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PMC:1540739 / 14131-21382 JSONTXT 5 Projects

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Id Subject Object Predicate Lexical cue
T11673 23-26 DT denotes The
T11674 27-36 NN denotes rationale
T11676 37-40 IN denotes for
T11677 41-45 NN denotes RMCE
T11679 45-46 HYPH denotes -
T11678 46-50 NN denotes ASAP
T11680 51-53 VBZ denotes is
T11675 54-62 VBN denotes detailed
T11681 63-65 IN denotes in
T11682 66-72 NN denotes Figure
T11683 73-74 CD denotes 1
T11684 74-75 . denotes .
T11685 75-228 sentence denotes The first step requires generating a floxed allele in ES cells that will serve as the substrate for subsequent exchanges (RMCE-ready ES cell, Figure 1).
T11686 76-79 DT denotes The
T11688 80-85 JJ denotes first
T11687 86-90 NN denotes step
T11689 91-99 VBZ denotes requires
T11690 100-110 VBG denotes generating
T11691 111-112 DT denotes a
T11693 113-119 VBN denotes floxed
T11692 120-126 NN denotes allele
T11694 127-129 IN denotes in
T11695 130-132 NN denotes ES
T11696 133-138 NNS denotes cells
T11697 139-143 WDT denotes that
T11699 144-148 MD denotes will
T11698 149-154 VB denotes serve
T11700 155-157 IN denotes as
T11701 158-161 DT denotes the
T11702 162-171 NN denotes substrate
T11703 172-175 IN denotes for
T11704 176-186 JJ denotes subsequent
T11705 187-196 NNS denotes exchanges
T11706 197-198 -LRB- denotes (
T11708 198-202 NN denotes RMCE
T11710 202-203 HYPH denotes -
T11709 203-208 JJ denotes ready
T11712 209-211 NN denotes ES
T11711 212-216 NN denotes cell
T11713 216-218 , denotes ,
T11707 218-224 NN denotes Figure
T11714 225-226 CD denotes 1
T11715 226-227 -RRB- denotes )
T11716 227-228 . denotes .
T11717 228-276 sentence denotes The targeting strategy is detailed in Figure 2.
T11718 229-232 DT denotes The
T11720 233-242 VBG denotes targeting
T11719 243-251 NN denotes strategy
T11722 252-254 VBZ denotes is
T11721 255-263 VBN denotes detailed
T11723 264-266 IN denotes in
T11724 267-273 NN denotes Figure
T11725 274-275 CD denotes 2
T11726 275-276 . denotes .
T11727 276-403 sentence denotes The frequency of targeting was 4% (12/300 puromycin-resistant clones, analyzed by Southern blot and long-range PCR, Figure 2).
T11728 277-280 DT denotes The
T11729 281-290 NN denotes frequency
T11731 291-293 IN denotes of
T11732 294-303 NN denotes targeting
T11730 304-307 VBD denotes was
T11733 308-309 CD denotes 4
T11734 309-310 NN denotes %
T11735 311-312 -LRB- denotes (
T11737 312-314 CD denotes 12
T11739 314-315 HYPH denotes /
T11738 315-318 CD denotes 300
T11741 319-328 NN denotes puromycin
T11743 328-329 HYPH denotes -
T11742 329-338 JJ denotes resistant
T11740 339-345 NNS denotes clones
T11744 345-347 , denotes ,
T11745 347-355 VBN denotes analyzed
T11746 356-358 IN denotes by
T11747 359-367 NNP denotes Southern
T11748 368-372 NN denotes blot
T11749 373-376 CC denotes and
T11750 377-381 JJ denotes long
T11752 381-382 HYPH denotes -
T11751 382-387 NN denotes range
T11753 388-391 NN denotes PCR
T11754 391-393 , denotes ,
T11736 393-399 NN denotes Figure
T11755 400-401 CD denotes 2
T11756 401-402 -RRB- denotes )
T11757 402-403 . denotes .
T11758 403-568 sentence denotes We next tested the efficiency of RMCE in ES cells, using a replacement construct encoding p53 fused to GFP (p53GFP) to enable tracking p53 in individual live cells.
T11759 404-406 PRP denotes We
T11761 407-411 RB denotes next
T11760 412-418 VBD denotes tested
T11762 419-422 DT denotes the
T11763 423-433 NN denotes efficiency
T11764 434-436 IN denotes of
T11765 437-441 NN denotes RMCE
T11766 442-444 IN denotes in
T11767 445-447 NN denotes ES
T11768 448-453 NNS denotes cells
T11769 453-455 , denotes ,
T11770 455-460 VBG denotes using
T11771 461-462 DT denotes a
T11773 463-474 NN denotes replacement
T11772 475-484 NN denotes construct
T11774 485-493 VBG denotes encoding
T11775 494-497 NN denotes p53
T11776 498-503 VBN denotes fused
T11777 504-506 IN denotes to
T11778 507-510 NN denotes GFP
T11779 511-512 -LRB- denotes (
T11780 512-518 NN denotes p53GFP
T11781 518-519 -RRB- denotes )
T11782 520-522 TO denotes to
T11783 523-529 VB denotes enable
T11784 530-538 VBG denotes tracking
T11785 539-542 NN denotes p53
T11786 543-545 IN denotes in
T11787 546-556 JJ denotes individual
T11789 557-561 JJ denotes live
T11788 562-567 NNS denotes cells
T11790 567-568 . denotes .
T11791 568-735 sentence denotes Importantly however, GFP fluorescence was not used to screen cells with targeted events, as we wanted to develop a general method to isolate marker-free recombinants.
T11792 569-580 RB denotes Importantly
T11794 581-588 RB denotes however
T11795 588-590 , denotes ,
T11796 590-593 NN denotes GFP
T11797 594-606 NN denotes fluorescence
T11798 607-610 VBD denotes was
T11799 611-614 RB denotes not
T11793 615-619 VBN denotes used
T11800 620-622 TO denotes to
T11801 623-629 VB denotes screen
T11802 630-635 NNS denotes cells
T11803 636-640 IN denotes with
T11804 641-649 VBN denotes targeted
T11805 650-656 NNS denotes events
T11806 656-658 , denotes ,
T11807 658-660 IN denotes as
T11809 661-663 PRP denotes we
T11808 664-670 VBD denotes wanted
T11810 671-673 TO denotes to
T11811 674-681 VB denotes develop
T11812 682-683 DT denotes a
T11814 684-691 JJ denotes general
T11813 692-698 NN denotes method
T11815 699-701 TO denotes to
T11816 702-709 VB denotes isolate
T11817 710-716 NN denotes marker
T11819 716-717 HYPH denotes -
T11818 717-721 JJ denotes free
T11820 722-734 NNS denotes recombinants
T11821 734-735 . denotes .
T11822 735-783 sentence denotes The exchange strategy is detailed in Figure 3A.
T11823 736-739 DT denotes The
T11825 740-748 NN denotes exchange
T11824 749-757 NN denotes strategy
T11827 758-760 VBZ denotes is
T11826 761-769 VBN denotes detailed
T11828 770-772 IN denotes in
T11829 773-779 NN denotes Figure
T11830 780-782 NN denotes 3A
T11831 782-783 . denotes .
T11832 783-954 sentence denotes We picked 65 ES cell clones resistant to 1-(-2-deoxy-2-fluoro-1-b-d-arabino-furanosyl)-5-iodouracil (FIAU) due to TK loss and analyzed their DNA by PCR and Southern blot.
T11833 784-786 PRP denotes We
T11834 787-793 VBD denotes picked
T11835 794-796 CD denotes 65
T11837 797-799 NN denotes ES
T11838 800-804 NN denotes cell
T11836 805-811 NNS denotes clones
T11839 812-821 JJ denotes resistant
T11840 822-824 IN denotes to
T11841 825-826 CD denotes 1
T11843 826-827 HYPH denotes -
T11844 827-828 -LRB- denotes (
T11846 828-829 HYPH denotes -
T11847 829-830 CD denotes 2
T11848 830-831 HYPH denotes -
T11849 831-836 NN denotes deoxy
T11850 836-837 HYPH denotes -
T11851 837-838 CD denotes 2
T11852 838-839 HYPH denotes -
T11853 839-845 NN denotes fluoro
T11854 845-846 HYPH denotes -
T11855 846-847 CD denotes 1
T11856 847-848 HYPH denotes -
T11857 848-849 NN denotes b
T11858 849-850 HYPH denotes -
T11859 850-851 NN denotes d
T11860 851-852 HYPH denotes -
T11861 852-859 NN denotes arabino
T11862 859-860 HYPH denotes -
T11845 860-869 NN denotes furanosyl
T11863 869-870 -RRB- denotes )
T11864 870-871 HYPH denotes -
T11842 871-872 CD denotes 5
T11866 872-873 HYPH denotes -
T11865 873-883 NN denotes iodouracil
T11867 884-885 -LRB- denotes (
T11868 885-889 NN denotes FIAU
T11869 889-890 -RRB- denotes )
T11870 891-894 IN denotes due
T11871 895-897 IN denotes to
T11872 898-900 NN denotes TK
T11873 901-905 NN denotes loss
T11874 906-909 CC denotes and
T11875 910-918 VBD denotes analyzed
T11876 919-924 PRP$ denotes their
T11877 925-928 NN denotes DNA
T11878 929-931 IN denotes by
T11879 932-935 NN denotes PCR
T11880 936-939 CC denotes and
T11881 940-948 NNP denotes Southern
T11882 949-953 NN denotes blot
T11883 953-954 . denotes .
T11884 954-1050 sentence denotes Strikingly, 54 proper recombinants were identified, indicating very high RMCE efficiency (83%).
T11885 955-965 RB denotes Strikingly
T11887 965-967 , denotes ,
T11888 967-969 CD denotes 54
T11890 970-976 JJ denotes proper
T11889 977-989 NNS denotes recombinants
T11891 990-994 VBD denotes were
T11886 995-1005 VBN denotes identified
T11892 1005-1007 , denotes ,
T11893 1007-1017 VBG denotes indicating
T11894 1018-1022 RB denotes very
T11895 1023-1027 JJ denotes high
T11897 1028-1032 NN denotes RMCE
T11896 1033-1043 NN denotes efficiency
T11898 1044-1045 -LRB- denotes (
T11900 1045-1047 CD denotes 83
T11899 1047-1048 NN denotes %
T11901 1048-1049 -RRB- denotes )
T11902 1049-1050 . denotes .
T11903 1050-1156 sentence denotes RMCE also proved to be precise, as no aberrant bands were detected in PCR and Southern blots (Figure 3A).
T11904 1051-1055 NN denotes RMCE
T11906 1056-1060 RB denotes also
T11905 1061-1067 VBD denotes proved
T11907 1068-1070 TO denotes to
T11908 1071-1073 VB denotes be
T11909 1074-1081 JJ denotes precise
T11910 1081-1083 , denotes ,
T11911 1083-1085 IN denotes as
T11913 1086-1088 DT denotes no
T11915 1089-1097 JJ denotes aberrant
T11914 1098-1103 NNS denotes bands
T11916 1104-1108 VBD denotes were
T11912 1109-1117 VBN denotes detected
T11917 1118-1120 IN denotes in
T11918 1121-1124 NN denotes PCR
T11919 1125-1128 CC denotes and
T11920 1129-1137 NNP denotes Southern
T11921 1138-1143 NNS denotes blots
T11922 1144-1145 -LRB- denotes (
T11924 1145-1151 NN denotes Figure
T11923 1152-1154 NN denotes 3A
T11925 1154-1155 -RRB- denotes )
T11926 1155-1156 . denotes .
T11927 1156-1310 sentence denotes p53+/GFP ES clones were analyzed by western blot with an antibody against p53, and found to express an additional band at the expected size (ca. 80 kDa).
T11928 1157-1160 NN denotes p53
T11930 1160-1161 SYM denotes +
T11931 1161-1162 HYPH denotes /
T11929 1162-1165 NN denotes GFP
T11933 1166-1168 NN denotes ES
T11932 1169-1175 NNS denotes clones
T11935 1176-1180 VBD denotes were
T11934 1181-1189 VBN denotes analyzed
T11936 1190-1192 IN denotes by
T11937 1193-1200 NNP denotes western
T11938 1201-1205 NN denotes blot
T11939 1206-1210 IN denotes with
T11940 1211-1213 DT denotes an
T11941 1214-1222 NN denotes antibody
T11942 1223-1230 IN denotes against
T11943 1231-1234 NN denotes p53
T11944 1234-1236 , denotes ,
T11945 1236-1239 CC denotes and
T11946 1240-1245 VBN denotes found
T11947 1246-1248 TO denotes to
T11948 1249-1256 VB denotes express
T11949 1257-1259 DT denotes an
T11951 1260-1270 JJ denotes additional
T11950 1271-1275 NN denotes band
T11952 1276-1278 IN denotes at
T11953 1279-1282 DT denotes the
T11955 1283-1291 VBN denotes expected
T11954 1292-1296 NN denotes size
T11956 1297-1298 -LRB- denotes (
T11957 1298-1301 NN denotes ca.
T11958 1302-1304 CD denotes 80
T11959 1305-1308 NN denotes kDa
T11960 1308-1309 -RRB- denotes )
T11961 1309-1310 . denotes .
T11962 1310-1634 sentence denotes Surprisingly, the fusion of GFP to p53 apparently altered p53 stability: steady-state levels of p53GFP were much higher than those of wild-type p53 (p53WT) in unstressed cells, and did not vary significantly after DNA damage, so that the levels for both p53WT and p53GFP were similar after adriamycin treatment (Figure 3A).
T11963 1311-1323 RB denotes Surprisingly
T11965 1323-1325 , denotes ,
T11966 1325-1328 DT denotes the
T11967 1329-1335 NN denotes fusion
T11968 1336-1338 IN denotes of
T11969 1339-1342 NN denotes GFP
T11970 1343-1345 IN denotes to
T11971 1346-1349 NN denotes p53
T11972 1350-1360 RB denotes apparently
T11964 1361-1368 VBD denotes altered
T11974 1369-1372 NN denotes p53
T11975 1373-1382 NN denotes stability
T11976 1382-1384 : denotes :
T11977 1384-1390 JJ denotes steady
T11979 1390-1391 HYPH denotes -
T11978 1391-1396 NN denotes state
T11980 1397-1403 NNS denotes levels
T11981 1404-1406 IN denotes of
T11982 1407-1413 NN denotes p53GFP
T11973 1414-1418 VBD denotes were
T11983 1419-1423 RB denotes much
T11984 1424-1430 JJR denotes higher
T11985 1431-1435 IN denotes than
T11986 1436-1441 DT denotes those
T11987 1442-1444 IN denotes of
T11988 1445-1449 JJ denotes wild
T11990 1449-1450 HYPH denotes -
T11989 1450-1454 NN denotes type
T11991 1455-1458 NN denotes p53
T11992 1459-1460 -LRB- denotes (
T11993 1460-1465 NN denotes p53WT
T11994 1465-1466 -RRB- denotes )
T11995 1467-1469 IN denotes in
T11996 1470-1480 JJ denotes unstressed
T11997 1481-1486 NNS denotes cells
T11998 1486-1488 , denotes ,
T11999 1488-1491 CC denotes and
T12000 1492-1495 VBD denotes did
T12002 1496-1499 RB denotes not
T12001 1500-1504 VB denotes vary
T12003 1505-1518 RB denotes significantly
T12004 1519-1524 IN denotes after
T12005 1525-1528 NN denotes DNA
T12006 1529-1535 NN denotes damage
T12007 1535-1537 , denotes ,
T12008 1537-1539 IN denotes so
T12010 1540-1544 IN denotes that
T12011 1545-1548 DT denotes the
T12012 1549-1555 NNS denotes levels
T12013 1556-1559 IN denotes for
T12014 1560-1564 CC denotes both
T12015 1565-1570 NN denotes p53WT
T12016 1571-1574 CC denotes and
T12017 1575-1581 NN denotes p53GFP
T12009 1582-1586 VBD denotes were
T12018 1587-1594 JJ denotes similar
T12019 1595-1600 IN denotes after
T12020 1601-1611 NN denotes adriamycin
T12021 1612-1621 NN denotes treatment
T12022 1622-1623 -LRB- denotes (
T12024 1623-1629 NN denotes Figure
T12023 1630-1632 NN denotes 3A
T12025 1632-1633 -RRB- denotes )
T12026 1633-1634 . denotes .
T12027 1634-1767 sentence denotes Six independent p53+/GFP clones were injected into blastocysts and transferred to pseudo-pregnant females using standard procedures.
T12028 1635-1638 CD denotes Six
T12030 1639-1650 JJ denotes independent
T12031 1651-1654 NN denotes p53
T12033 1654-1655 SYM denotes +
T12034 1655-1656 HYPH denotes /
T12032 1656-1659 NN denotes GFP
T12029 1660-1666 NNS denotes clones
T12036 1667-1671 VBD denotes were
T12035 1672-1680 VBN denotes injected
T12037 1681-1685 IN denotes into
T12038 1686-1697 NNS denotes blastocysts
T12039 1698-1701 CC denotes and
T12040 1702-1713 VBN denotes transferred
T12041 1714-1716 IN denotes to
T12042 1717-1723 JJ denotes pseudo
T12044 1723-1724 HYPH denotes -
T12043 1724-1732 JJ denotes pregnant
T12045 1733-1740 NNS denotes females
T12046 1741-1746 VBG denotes using
T12047 1747-1755 JJ denotes standard
T12048 1756-1766 NNS denotes procedures
T12049 1766-1767 . denotes .
T12050 1767-1809 sentence denotes Strikingly, no pregnancies were obtained.
T12051 1768-1778 RB denotes Strikingly
T12053 1778-1780 , denotes ,
T12054 1780-1782 DT denotes no
T12055 1783-1794 NNS denotes pregnancies
T12056 1795-1799 VBD denotes were
T12052 1800-1808 VBN denotes obtained
T12057 1808-1809 . denotes .
T12058 1809-2023 sentence denotes It has been shown that the p53 pathway is regulated very differently in ES and somatic cells: ES cells contain relatively high p53 levels and lack the p53-mediated DNA damage responses found in somatic cells (18).
T12059 1810-1812 PRP denotes It
T12061 1813-1816 VBZ denotes has
T12062 1817-1821 VBN denotes been
T12060 1822-1827 VBN denotes shown
T12064 1828-1832 IN denotes that
T12066 1833-1836 DT denotes the
T12068 1837-1840 NN denotes p53
T12067 1841-1848 NN denotes pathway
T12069 1849-1851 VBZ denotes is
T12065 1852-1861 VBN denotes regulated
T12070 1862-1866 RB denotes very
T12071 1867-1878 RB denotes differently
T12072 1879-1881 IN denotes in
T12073 1882-1884 NN denotes ES
T12075 1885-1888 CC denotes and
T12076 1889-1896 JJ denotes somatic
T12074 1897-1902 NNS denotes cells
T12077 1902-1904 : denotes :
T12078 1904-1906 NN denotes ES
T12079 1907-1912 NNS denotes cells
T12063 1913-1920 VBP denotes contain
T12080 1921-1931 RB denotes relatively
T12081 1932-1936 JJ denotes high
T12083 1937-1940 NN denotes p53
T12082 1941-1947 NNS denotes levels
T12084 1948-1951 CC denotes and
T12085 1952-1956 VBP denotes lack
T12086 1957-1960 DT denotes the
T12088 1961-1964 NN denotes p53
T12089 1964-1965 HYPH denotes -
T12090 1965-1973 JJ denotes mediated
T12091 1974-1977 NN denotes DNA
T12092 1978-1984 NN denotes damage
T12087 1985-1994 NNS denotes responses
T12093 1995-2000 VBN denotes found
T12094 2001-2003 IN denotes in
T12095 2004-2011 JJ denotes somatic
T12096 2012-2017 NNS denotes cells
T12097 2018-2019 -LRB- denotes (
T12098 2019-2021 CD denotes 18
T12099 2021-2022 -RRB- denotes )
T12100 2022-2023 . denotes .
T12101 2023-2395 sentence denotes This, together with the observation that p53 levels decrease during mouse embryogenesis (19), suggested an explanation for the observed lack of pregnancies: we speculate that the high levels of p53GFP in the ES cells injected into blastocysts might have prevented normal embryonic development once these cells began to differentiate and the p53 pathway became functional.
T12102 2024-2028 DT denotes This
T12104 2028-2030 , denotes ,
T12105 2030-2038 RB denotes together
T12106 2039-2043 IN denotes with
T12107 2044-2047 DT denotes the
T12108 2048-2059 NN denotes observation
T12109 2060-2064 IN denotes that
T12111 2065-2068 NN denotes p53
T12112 2069-2075 NNS denotes levels
T12110 2076-2084 VBP denotes decrease
T12113 2085-2091 IN denotes during
T12114 2092-2097 NN denotes mouse
T12115 2098-2111 NN denotes embryogenesis
T12116 2112-2113 -LRB- denotes (
T12117 2113-2115 CD denotes 19
T12118 2115-2116 -RRB- denotes )
T12119 2116-2118 , denotes ,
T12103 2118-2127 VBD denotes suggested
T12121 2128-2130 DT denotes an
T12122 2131-2142 NN denotes explanation
T12123 2143-2146 IN denotes for
T12124 2147-2150 DT denotes the
T12126 2151-2159 VBN denotes observed
T12125 2160-2164 NN denotes lack
T12127 2165-2167 IN denotes of
T12128 2168-2179 NNS denotes pregnancies
T12129 2179-2181 : denotes :
T12130 2181-2183 PRP denotes we
T12120 2184-2193 VBP denotes speculate
T12131 2194-2198 IN denotes that
T12133 2199-2202 DT denotes the
T12135 2203-2207 JJ denotes high
T12134 2208-2214 NNS denotes levels
T12136 2215-2217 IN denotes of
T12137 2218-2224 NN denotes p53GFP
T12138 2225-2227 IN denotes in
T12139 2228-2231 DT denotes the
T12141 2232-2234 NN denotes ES
T12140 2235-2240 NNS denotes cells
T12142 2241-2249 VBN denotes injected
T12143 2250-2254 IN denotes into
T12144 2255-2266 NNS denotes blastocysts
T12145 2267-2272 MD denotes might
T12146 2273-2277 VB denotes have
T12132 2278-2287 VBN denotes prevented
T12147 2288-2294 JJ denotes normal
T12149 2295-2304 JJ denotes embryonic
T12148 2305-2316 NN denotes development
T12150 2317-2321 IN denotes once
T12152 2322-2327 DT denotes these
T12153 2328-2333 NNS denotes cells
T12151 2334-2339 VBD denotes began
T12154 2340-2342 TO denotes to
T12155 2343-2356 VB denotes differentiate
T12156 2357-2360 CC denotes and
T12157 2361-2364 DT denotes the
T12159 2365-2368 NN denotes p53
T12158 2369-2376 NN denotes pathway
T12160 2377-2383 VBD denotes became
T12161 2384-2394 JJ denotes functional
T12162 2394-2395 . denotes .
T12163 2395-2558 sentence denotes To test this possibility, we performed RMCE with a p53 fusion gene in which the p53 proline-rich domain (PRD) was deleted (p53ΔP), and the p53ΔP was fused to GFP.
T12164 2396-2398 TO denotes To
T12165 2399-2403 VB denotes test
T12167 2404-2408 DT denotes this
T12168 2409-2420 NN denotes possibility
T12169 2420-2422 , denotes ,
T12170 2422-2424 PRP denotes we
T12166 2425-2434 VBD denotes performed
T12171 2435-2439 NN denotes RMCE
T12172 2440-2444 IN denotes with
T12173 2445-2446 DT denotes a
T12175 2447-2450 NN denotes p53
T12176 2451-2457 NN denotes fusion
T12174 2458-2462 NN denotes gene
T12177 2463-2465 IN denotes in
T12179 2466-2471 WDT denotes which
T12180 2472-2475 DT denotes the
T12182 2476-2479 NN denotes p53
T12183 2480-2487 NN denotes proline
T12185 2487-2488 HYPH denotes -
T12184 2488-2492 JJ denotes rich
T12181 2493-2499 NN denotes domain
T12186 2500-2501 -LRB- denotes (
T12187 2501-2504 NN denotes PRD
T12188 2504-2505 -RRB- denotes )
T12189 2506-2509 VBD denotes was
T12178 2510-2517 VBN denotes deleted
T12190 2518-2519 -LRB- denotes (
T12191 2519-2524 NN denotes p53ΔP
T12192 2524-2525 -RRB- denotes )
T12193 2525-2527 , denotes ,
T12194 2527-2530 CC denotes and
T12195 2531-2534 DT denotes the
T12196 2535-2540 NN denotes p53ΔP
T12198 2541-2544 VBD denotes was
T12197 2545-2550 VBN denotes fused
T12199 2551-2553 IN denotes to
T12200 2554-2557 NN denotes GFP
T12201 2557-2558 . denotes .
T12202 2558-2689 sentence denotes We used this mutant because deleting the proline-rich domain decreases stability and compromises DNA-damage responses in vivo (5).
T12203 2559-2561 PRP denotes We
T12204 2562-2566 VBD denotes used
T12205 2567-2571 DT denotes this
T12206 2572-2578 NN denotes mutant
T12207 2579-2586 IN denotes because
T12209 2587-2595 VBG denotes deleting
T12210 2596-2599 DT denotes the
T12212 2600-2607 NN denotes proline
T12214 2607-2608 HYPH denotes -
T12213 2608-2612 JJ denotes rich
T12211 2613-2619 NN denotes domain
T12208 2620-2629 VBZ denotes decreases
T12215 2630-2639 NN denotes stability
T12216 2640-2643 CC denotes and
T12217 2644-2655 VBZ denotes compromises
T12218 2656-2659 NN denotes DNA
T12220 2659-2660 HYPH denotes -
T12219 2660-2666 NN denotes damage
T12221 2667-2676 NNS denotes responses
T12222 2677-2679 FW denotes in
T12223 2680-2684 FW denotes vivo
T12224 2685-2686 -LRB- denotes (
T12225 2686-2687 CD denotes 5
T12226 2687-2688 -RRB- denotes )
T12227 2688-2689 . denotes .
T12228 2689-2784 sentence denotes According to our hypothesis, this hypomorphic mutant should not prevent embryonic development.
T12229 2690-2699 VBG denotes According
T12231 2700-2702 IN denotes to
T12232 2703-2706 PRP$ denotes our
T12233 2707-2717 NN denotes hypothesis
T12234 2717-2719 , denotes ,
T12235 2719-2723 DT denotes this
T12237 2724-2735 JJ denotes hypomorphic
T12236 2736-2742 NN denotes mutant
T12238 2743-2749 MD denotes should
T12239 2750-2753 RB denotes not
T12230 2754-2761 VB denotes prevent
T12240 2762-2771 JJ denotes embryonic
T12241 2772-2783 NN denotes development
T12242 2783-2784 . denotes .
T12243 2784-2971 sentence denotes RMCE with a p53ΔPGFP replacement plasmid was again very efficient and western blots revealed an additional band of the predicted molecular weight only in p53+/ΔPGFP ES cells (Figure 3B).
T12244 2785-2789 NN denotes RMCE
T12246 2790-2794 IN denotes with
T12247 2795-2796 DT denotes a
T12249 2797-2805 NN denotes p53ΔPGFP
T12250 2806-2817 NN denotes replacement
T12248 2818-2825 NN denotes plasmid
T12245 2826-2829 VBD denotes was
T12251 2830-2835 RB denotes again
T12252 2836-2840 RB denotes very
T12253 2841-2850 JJ denotes efficient
T12254 2851-2854 CC denotes and
T12255 2855-2862 NNP denotes western
T12256 2863-2868 NNS denotes blots
T12257 2869-2877 VBD denotes revealed
T12258 2878-2880 DT denotes an
T12260 2881-2891 JJ denotes additional
T12259 2892-2896 NN denotes band
T12261 2897-2899 IN denotes of
T12262 2900-2903 DT denotes the
T12264 2904-2913 VBN denotes predicted
T12265 2914-2923 JJ denotes molecular
T12263 2924-2930 NN denotes weight
T12266 2931-2935 RB denotes only
T12267 2936-2938 IN denotes in
T12268 2939-2942 NN denotes p53
T12270 2942-2943 SYM denotes +
T12271 2943-2944 HYPH denotes /
T12269 2944-2949 NN denotes ΔPGFP
T12273 2950-2952 NN denotes ES
T12272 2953-2958 NNS denotes cells
T12274 2959-2960 -LRB- denotes (
T12276 2960-2966 NN denotes Figure
T12275 2967-2969 NN denotes 3B
T12277 2969-2970 -RRB- denotes )
T12278 2970-2971 . denotes .
T12279 2971-3123 sentence denotes As expected, the PRD deletion correlated with lower expression levels: p53ΔPGFP was much less abundant than p53WT in all p53+/ΔPGFP clones (Figure 3B).
T12280 2972-2974 IN denotes As
T12281 2975-2983 VBN denotes expected
T12283 2983-2985 , denotes ,
T12284 2985-2988 DT denotes the
T12286 2989-2992 NN denotes PRD
T12285 2993-3001 NN denotes deletion
T12282 3002-3012 VBD denotes correlated
T12288 3013-3017 IN denotes with
T12289 3018-3023 JJR denotes lower
T12291 3024-3034 NN denotes expression
T12290 3035-3041 NNS denotes levels
T12292 3041-3043 : denotes :
T12293 3043-3051 NN denotes p53ΔPGFP
T12287 3052-3055 VBD denotes was
T12294 3056-3060 RB denotes much
T12295 3061-3065 RBR denotes less
T12296 3066-3074 JJ denotes abundant
T12297 3075-3079 IN denotes than
T12298 3080-3085 NN denotes p53WT
T12299 3086-3088 IN denotes in
T12300 3089-3092 DT denotes all
T12302 3093-3096 NN denotes p53
T12304 3096-3097 SYM denotes +
T12305 3097-3098 HYPH denotes /
T12303 3098-3103 NN denotes ΔPGFP
T12301 3104-3110 NNS denotes clones
T12306 3111-3112 -LRB- denotes (
T12308 3112-3118 NN denotes Figure
T12307 3119-3121 NN denotes 3B
T12309 3121-3122 -RRB- denotes )
T12310 3122-3123 . denotes .
T12311 3123-3254 sentence denotes We next determined whether p53+/ΔPGFP ES cells could generate chimeric mice and transmit the modified allele through the germline.
T12312 3124-3126 PRP denotes We
T12314 3127-3131 RB denotes next
T12313 3132-3142 VBD denotes determined
T12315 3143-3150 IN denotes whether
T12317 3151-3154 NN denotes p53
T12319 3154-3155 SYM denotes +
T12320 3155-3156 HYPH denotes /
T12318 3156-3161 NN denotes ΔPGFP
T12322 3162-3164 NN denotes ES
T12321 3165-3170 NNS denotes cells
T12323 3171-3176 MD denotes could
T12316 3177-3185 VB denotes generate
T12324 3186-3194 JJ denotes chimeric
T12325 3195-3199 NNS denotes mice
T12326 3200-3203 CC denotes and
T12327 3204-3212 VB denotes transmit
T12328 3213-3216 DT denotes the
T12330 3217-3225 VBN denotes modified
T12329 3226-3232 NN denotes allele
T12331 3233-3240 IN denotes through
T12332 3241-3244 DT denotes the
T12333 3245-3253 NN denotes germline
T12334 3253-3254 . denotes .
T12335 3254-3362 sentence denotes Two p53+/ΔPGFP ES cell clones were injected into blastocysts and highly chimeric (>80%) mice were obtained.
T12336 3255-3258 CD denotes Two
T12338 3259-3262 NN denotes p53
T12340 3262-3263 SYM denotes +
T12341 3263-3264 HYPH denotes /
T12339 3264-3269 NN denotes ΔPGFP
T12342 3270-3272 NN denotes ES
T12343 3273-3277 NN denotes cell
T12337 3278-3284 NNS denotes clones
T12345 3285-3289 VBD denotes were
T12344 3290-3298 VBN denotes injected
T12346 3299-3303 IN denotes into
T12347 3304-3315 NNS denotes blastocysts
T12348 3316-3319 CC denotes and
T12349 3320-3326 RB denotes highly
T12350 3327-3335 JJ denotes chimeric
T12352 3336-3337 -LRB- denotes (
T12353 3337-3338 SYM denotes >
T12354 3338-3340 CD denotes 80
T12351 3340-3341 NN denotes %
T12356 3341-3342 -RRB- denotes )
T12355 3343-3347 NNS denotes mice
T12358 3348-3352 VBD denotes were
T12357 3353-3361 VBN denotes obtained
T12359 3361-3362 . denotes .
T12360 3362-3450 sentence denotes Heterozygote pups were recovered from the mating the chimeras with WT mice (Figure 3C).
T12361 3363-3375 NN denotes Heterozygote
T12362 3376-3380 NNS denotes pups
T12364 3381-3385 VBD denotes were
T12363 3386-3395 VBN denotes recovered
T12365 3396-3400 IN denotes from
T12366 3401-3404 DT denotes the
T12367 3405-3411 NN denotes mating
T12368 3412-3415 DT denotes the
T12369 3416-3424 NNS denotes chimeras
T12370 3425-3429 IN denotes with
T12371 3430-3432 NN denotes WT
T12372 3433-3437 NNS denotes mice
T12373 3438-3439 -LRB- denotes (
T12375 3439-3445 NN denotes Figure
T12374 3446-3448 NN denotes 3C
T12376 3448-3449 -RRB- denotes )
T12377 3449-3450 . denotes .
T12378 3450-3605 sentence denotes These data demonstrate that marker-free RMCE is very efficient in ES cells and allows germline transmission of a targeted mutation (see Figure 1, path A).
T12379 3451-3456 DT denotes These
T12380 3457-3461 NNS denotes data
T12381 3462-3473 VBP denotes demonstrate
T12382 3474-3478 IN denotes that
T12384 3479-3485 NN denotes marker
T12386 3485-3486 HYPH denotes -
T12385 3486-3490 JJ denotes free
T12387 3491-3495 NN denotes RMCE
T12383 3496-3498 VBZ denotes is
T12388 3499-3503 RB denotes very
T12389 3504-3513 JJ denotes efficient
T12390 3514-3516 IN denotes in
T12391 3517-3519 NN denotes ES
T12392 3520-3525 NNS denotes cells
T12393 3526-3529 CC denotes and
T12394 3530-3536 VBZ denotes allows
T12395 3537-3545 NN denotes germline
T12396 3546-3558 NN denotes transmission
T12397 3559-3561 IN denotes of
T12398 3562-3563 DT denotes a
T12400 3564-3572 VBN denotes targeted
T12399 3573-3581 NN denotes mutation
T12401 3582-3583 -LRB- denotes (
T12402 3583-3586 VB denotes see
T12403 3587-3593 NN denotes Figure
T12404 3594-3595 CD denotes 1
T12405 3595-3597 , denotes ,
T12406 3597-3601 NN denotes path
T12407 3602-3603 NN denotes A
T12408 3603-3604 -RRB- denotes )
T12409 3604-3605 . denotes .
T12410 3605-3739 sentence denotes We next determined whether the RMCE approach could be used to target mutations at the p53 allele in somatic cells (Figure 1, path B).
T12411 3606-3608 PRP denotes We
T12413 3609-3613 RB denotes next
T12412 3614-3624 VBD denotes determined
T12414 3625-3632 IN denotes whether
T12416 3633-3636 DT denotes the
T12418 3637-3641 NN denotes RMCE
T12417 3642-3650 NN denotes approach
T12419 3651-3656 MD denotes could
T12420 3657-3659 VB denotes be
T12415 3660-3664 VBN denotes used
T12421 3665-3667 TO denotes to
T12422 3668-3674 VB denotes target
T12423 3675-3684 NNS denotes mutations
T12424 3685-3687 IN denotes at
T12425 3688-3691 DT denotes the
T12427 3692-3695 NN denotes p53
T12426 3696-3702 NN denotes allele
T12428 3703-3705 IN denotes in
T12429 3706-3713 JJ denotes somatic
T12430 3714-3719 NNS denotes cells
T12431 3720-3721 -LRB- denotes (
T12432 3721-3727 NN denotes Figure
T12433 3728-3729 CD denotes 1
T12434 3729-3731 , denotes ,
T12435 3731-3735 NN denotes path
T12436 3736-3737 NN denotes B
T12437 3737-3738 -RRB- denotes )
T12438 3738-3739 . denotes .
T12439 3739-3902 sentence denotes We first verified that the RMCE-ready p53 locus (p53RMCE) could be transmitted through the germline by mating p53RMCE/+ chimeras with p53+/− mice (20) (Figure 4).
T12440 3740-3742 PRP denotes We
T12442 3743-3748 RB denotes first
T12441 3749-3757 VBD denotes verified
T12443 3758-3762 IN denotes that
T12445 3763-3766 DT denotes the
T12447 3767-3771 NN denotes RMCE
T12449 3771-3772 HYPH denotes -
T12448 3772-3777 JJ denotes ready
T12450 3778-3781 NN denotes p53
T12446 3782-3787 NN denotes locus
T12451 3788-3789 -LRB- denotes (
T12452 3789-3796 NN denotes p53RMCE
T12453 3796-3797 -RRB- denotes )
T12454 3798-3803 MD denotes could
T12455 3804-3806 VB denotes be
T12444 3807-3818 VBN denotes transmitted
T12456 3819-3826 IN denotes through
T12457 3827-3830 DT denotes the
T12458 3831-3839 NN denotes germline
T12459 3840-3842 IN denotes by
T12460 3843-3849 VBG denotes mating
T12461 3850-3857 NN denotes p53RMCE
T12463 3857-3858 HYPH denotes /
T12464 3858-3859 SYM denotes +
T12462 3860-3868 NNS denotes chimeras
T12465 3869-3873 IN denotes with
T12466 3874-3877 NN denotes p53
T12468 3877-3878 SYM denotes +
T12469 3878-3879 HYPH denotes /
T12470 3879-3880 SYM denotes −
T12467 3881-3885 NNS denotes mice
T12471 3886-3887 -LRB- denotes (
T12472 3887-3889 CD denotes 20
T12473 3889-3890 -RRB- denotes )
T12474 3891-3892 -LRB- denotes (
T12475 3892-3898 NN denotes Figure
T12476 3899-3900 CD denotes 4
T12477 3900-3901 -RRB- denotes )
T12478 3901-3902 . denotes .
T12479 3902-4023 sentence denotes Importantly, this allowed us to generate p53RMCE/− MEFs, which were used to test RMCE at the p53 locus in somatic cells.
T12480 3903-3914 RB denotes Importantly
T12482 3914-3916 , denotes ,
T12483 3916-3920 DT denotes this
T12481 3921-3928 VBD denotes allowed
T12484 3929-3931 PRP denotes us
T12486 3932-3934 TO denotes to
T12485 3935-3943 VB denotes generate
T12487 3944-3951 NN denotes p53RMCE
T12489 3951-3952 HYPH denotes /
T12490 3952-3953 SYM denotes −
T12488 3954-3958 NNS denotes MEFs
T12491 3958-3960 , denotes ,
T12492 3960-3965 WDT denotes which
T12494 3966-3970 VBD denotes were
T12493 3971-3975 VBN denotes used
T12495 3976-3978 TO denotes to
T12496 3979-3983 VB denotes test
T12497 3984-3988 NN denotes RMCE
T12498 3989-3991 IN denotes at
T12499 3992-3995 DT denotes the
T12501 3996-3999 NN denotes p53
T12500 4000-4005 NN denotes locus
T12502 4006-4008 IN denotes in
T12503 4009-4016 JJ denotes somatic
T12504 4017-4022 NNS denotes cells
T12505 4022-4023 . denotes .
T12506 4023-4191 sentence denotes We first attempted RMCE in MEFs by electroporating p53RMCE/− MEFs with a Cre-expression plasmid and the p53GFP plasmid, followed by selection with FIAU or ganciclovir.
T12507 4024-4026 PRP denotes We
T12509 4027-4032 RB denotes first
T12508 4033-4042 VBD denotes attempted
T12510 4043-4047 NN denotes RMCE
T12511 4048-4050 IN denotes in
T12512 4051-4055 NNS denotes MEFs
T12513 4056-4058 IN denotes by
T12514 4059-4074 VBG denotes electroporating
T12515 4075-4082 NN denotes p53RMCE
T12517 4082-4083 HYPH denotes /
T12518 4083-4084 SYM denotes −
T12516 4085-4089 NNS denotes MEFs
T12519 4090-4094 IN denotes with
T12520 4095-4096 DT denotes a
T12522 4097-4100 NN denotes Cre
T12524 4100-4101 HYPH denotes -
T12523 4101-4111 NN denotes expression
T12521 4112-4119 NN denotes plasmid
T12525 4120-4123 CC denotes and
T12526 4124-4127 DT denotes the
T12528 4128-4134 NN denotes p53GFP
T12527 4135-4142 NN denotes plasmid
T12529 4142-4144 , denotes ,
T12530 4144-4152 VBN denotes followed
T12531 4153-4155 IN denotes by
T12532 4156-4165 NN denotes selection
T12533 4166-4170 IN denotes with
T12534 4171-4175 NN denotes FIAU
T12535 4176-4178 CC denotes or
T12536 4179-4190 NN denotes ganciclovir
T12537 4190-4191 . denotes .
T12538 4191-4272 sentence denotes Strikingly, no clones with an exchanged allele were identified (data not shown).
T12539 4192-4202 RB denotes Strikingly
T12541 4202-4204 , denotes ,
T12542 4204-4206 DT denotes no
T12543 4207-4213 NNS denotes clones
T12544 4214-4218 IN denotes with
T12545 4219-4221 DT denotes an
T12547 4222-4231 VBN denotes exchanged
T12546 4232-4238 NN denotes allele
T12548 4239-4243 VBD denotes were
T12540 4244-4254 VBN denotes identified
T12549 4255-4256 -LRB- denotes (
T12551 4256-4260 NNS denotes data
T12552 4261-4264 RB denotes not
T12550 4265-4270 VBN denotes shown
T12553 4270-4271 -RRB- denotes )
T12554 4271-4272 . denotes .
T12555 4272-4475 sentence denotes RMCE with p53GFP in ES cells showed that p53GFP is expressed at high levels (Figure 3A), and as mentioned before, the p53 pathway that can be activated in MEFs is not readily activated in ES cells (18).
T12556 4273-4277 NN denotes RMCE
T12558 4278-4282 IN denotes with
T12559 4283-4289 NN denotes p53GFP
T12560 4290-4292 IN denotes in
T12561 4293-4295 NN denotes ES
T12562 4296-4301 NNS denotes cells
T12557 4302-4308 VBD denotes showed
T12563 4309-4313 IN denotes that
T12565 4314-4320 NN denotes p53GFP
T12566 4321-4323 VBZ denotes is
T12564 4324-4333 VBN denotes expressed
T12567 4334-4336 IN denotes at
T12568 4337-4341 JJ denotes high
T12569 4342-4348 NNS denotes levels
T12570 4349-4350 -LRB- denotes (
T12572 4350-4356 NN denotes Figure
T12571 4357-4359 NN denotes 3A
T12573 4359-4360 -RRB- denotes )
T12574 4360-4362 , denotes ,
T12575 4362-4365 CC denotes and
T12576 4366-4368 IN denotes as
T12577 4369-4378 VBN denotes mentioned
T12579 4379-4385 RB denotes before
T12580 4385-4387 , denotes ,
T12581 4387-4390 DT denotes the
T12583 4391-4394 NN denotes p53
T12582 4395-4402 NN denotes pathway
T12584 4403-4407 WDT denotes that
T12586 4408-4411 MD denotes can
T12587 4412-4414 VB denotes be
T12585 4415-4424 VBN denotes activated
T12588 4425-4427 IN denotes in
T12589 4428-4432 NNS denotes MEFs
T12590 4433-4435 VBZ denotes is
T12591 4436-4439 RB denotes not
T12592 4440-4447 RB denotes readily
T12578 4448-4457 VBN denotes activated
T12593 4458-4460 IN denotes in
T12594 4461-4463 NN denotes ES
T12595 4464-4469 NNS denotes cells
T12596 4470-4471 -LRB- denotes (
T12597 4471-4473 CD denotes 18
T12598 4473-4474 -RRB- denotes )
T12599 4474-4475 . denotes .
T12600 4475-4644 sentence denotes The results above suggest that high levels of p53GFP could be tolerated by ES cells but toxic to MEFs, so that MEFs in which an RMCE had occurred failed to proliferate.
T12601 4476-4479 DT denotes The
T12602 4480-4487 NNS denotes results
T12604 4488-4493 RB denotes above
T12603 4494-4501 VBP denotes suggest
T12605 4502-4506 IN denotes that
T12607 4507-4511 JJ denotes high
T12608 4512-4518 NNS denotes levels
T12609 4519-4521 IN denotes of
T12610 4522-4528 NN denotes p53GFP
T12611 4529-4534 MD denotes could
T12606 4535-4537 VB denotes be
T12612 4538-4547 VBN denotes tolerated
T12613 4548-4550 IN denotes by
T12614 4551-4553 NN denotes ES
T12615 4554-4559 NNS denotes cells
T12616 4560-4563 CC denotes but
T12617 4564-4569 JJ denotes toxic
T12618 4570-4572 IN denotes to
T12619 4573-4577 NNS denotes MEFs
T12620 4577-4579 , denotes ,
T12621 4579-4581 IN denotes so
T12623 4582-4586 IN denotes that
T12624 4587-4591 NNS denotes MEFs
T12625 4592-4594 IN denotes in
T12627 4595-4600 WDT denotes which
T12628 4601-4603 DT denotes an
T12629 4604-4608 NN denotes RMCE
T12630 4609-4612 VBD denotes had
T12626 4613-4621 VBN denotes occurred
T12622 4622-4628 VBD denotes failed
T12631 4629-4631 TO denotes to
T12632 4632-4643 VB denotes proliferate
T12633 4643-4644 . denotes .
T12634 4644-4854 sentence denotes To test this possibility, p53RMCE/− MEFs were electroporated with the p53GFP replacement construct with or without a Cre-expression plasmid, then analyzed by fluorescence microscopy 48 h after electroporation.
T12635 4645-4647 TO denotes To
T12636 4648-4652 VB denotes test
T12638 4653-4657 DT denotes this
T12639 4658-4669 NN denotes possibility
T12640 4669-4671 , denotes ,
T12641 4671-4678 NN denotes p53RMCE
T12643 4678-4679 HYPH denotes /
T12644 4679-4680 SYM denotes −
T12642 4681-4685 NNS denotes MEFs
T12645 4686-4690 VBD denotes were
T12637 4691-4705 VBN denotes electroporated
T12646 4706-4710 IN denotes with
T12647 4711-4714 DT denotes the
T12649 4715-4721 NN denotes p53GFP
T12650 4722-4733 NN denotes replacement
T12648 4734-4743 NN denotes construct
T12651 4744-4748 IN denotes with
T12652 4749-4751 CC denotes or
T12653 4752-4759 IN denotes without
T12654 4760-4761 DT denotes a
T12656 4762-4765 NN denotes Cre
T12658 4765-4766 HYPH denotes -
T12657 4766-4776 NN denotes expression
T12655 4777-4784 NN denotes plasmid
T12659 4784-4786 , denotes ,
T12660 4786-4790 RB denotes then
T12661 4791-4799 VBN denotes analyzed
T12662 4800-4802 IN denotes by
T12663 4803-4815 NN denotes fluorescence
T12664 4816-4826 NN denotes microscopy
T12665 4827-4829 CD denotes 48
T12666 4830-4831 NN denotes h
T12667 4832-4837 IN denotes after
T12668 4838-4853 NN denotes electroporation
T12669 4853-4854 . denotes .
T12670 4854-5072 sentence denotes The experiment was done without selection to enable observation of cells under conditions where a failure to proliferate would not derive from FIAU or ganciclovir toxicity but rather solely from the effects of p53GFP.
T12671 4855-4858 DT denotes The
T12672 4859-4869 NN denotes experiment
T12674 4870-4873 VBD denotes was
T12673 4874-4878 VBN denotes done
T12675 4879-4886 IN denotes without
T12676 4887-4896 NN denotes selection
T12677 4897-4899 TO denotes to
T12678 4900-4906 VB denotes enable
T12679 4907-4918 NN denotes observation
T12680 4919-4921 IN denotes of
T12681 4922-4927 NNS denotes cells
T12682 4928-4933 IN denotes under
T12683 4934-4944 NNS denotes conditions
T12684 4945-4950 WRB denotes where
T12686 4951-4952 DT denotes a
T12687 4953-4960 NN denotes failure
T12688 4961-4963 TO denotes to
T12689 4964-4975 VB denotes proliferate
T12690 4976-4981 MD denotes would
T12691 4982-4985 RB denotes not
T12685 4986-4992 VB denotes derive
T12692 4993-4997 IN denotes from
T12693 4998-5002 NN denotes FIAU
T12695 5003-5005 CC denotes or
T12696 5006-5017 NN denotes ganciclovir
T12694 5018-5026 NN denotes toxicity
T12697 5027-5030 CC denotes but
T12698 5031-5037 RB denotes rather
T12699 5038-5044 RB denotes solely
T12700 5045-5049 IN denotes from
T12701 5050-5053 DT denotes the
T12702 5054-5061 NNS denotes effects
T12703 5062-5064 IN denotes of
T12704 5065-5071 NN denotes p53GFP
T12705 5071-5072 . denotes .
T12706 5072-5215 sentence denotes We observed a few fluorescent cells only when the Cre expression plasmid was co-electroporated, suggesting that such cells resulted from RMCE.
T12707 5073-5075 PRP denotes We
T12708 5076-5084 VBD denotes observed
T12709 5085-5086 DT denotes a
T12711 5087-5090 JJ denotes few
T12712 5091-5102 JJ denotes fluorescent
T12710 5103-5108 NNS denotes cells
T12713 5109-5113 RB denotes only
T12714 5114-5118 WRB denotes when
T12716 5119-5122 DT denotes the
T12718 5123-5126 NN denotes Cre
T12719 5127-5137 NN denotes expression
T12717 5138-5145 NN denotes plasmid
T12720 5146-5149 VBD denotes was
T12715 5150-5167 VBN denotes co-electroporated
T12721 5167-5169 , denotes ,
T12722 5169-5179 VBG denotes suggesting
T12723 5180-5184 IN denotes that
T12725 5185-5189 JJ denotes such
T12726 5190-5195 NNS denotes cells
T12724 5196-5204 VBD denotes resulted
T12727 5205-5209 IN denotes from
T12728 5210-5214 NN denotes RMCE
T12729 5214-5215 . denotes .
T12730 5215-5398 sentence denotes Importantly, the rare fluorescent cells had a flat, ‘fried-egg’ appearance typical of senescent cells (Figure 5A), and when plates were observed 5 days later, the cells had detached.
T12731 5216-5227 RB denotes Importantly
T12733 5227-5229 , denotes ,
T12734 5229-5232 DT denotes the
T12736 5233-5237 JJ denotes rare
T12737 5238-5249 JJ denotes fluorescent
T12735 5250-5255 NNS denotes cells
T12732 5256-5259 VBD denotes had
T12738 5260-5261 DT denotes a
T12740 5262-5266 JJ denotes flat
T12741 5266-5268 , denotes ,
T12742 5268-5269 `` denotes ‘
T12743 5269-5274 JJ denotes fried
T12745 5274-5275 HYPH denotes -
T12744 5275-5278 NN denotes egg
T12746 5278-5279 '' denotes ’
T12739 5280-5290 NN denotes appearance
T12747 5291-5298 JJ denotes typical
T12748 5299-5301 IN denotes of
T12749 5302-5311 JJ denotes senescent
T12750 5312-5317 NNS denotes cells
T12751 5318-5319 -LRB- denotes (
T12753 5319-5325 NN denotes Figure
T12752 5326-5328 NN denotes 5A
T12754 5328-5329 -RRB- denotes )
T12755 5329-5331 , denotes ,
T12756 5331-5334 CC denotes and
T12757 5335-5339 WRB denotes when
T12759 5340-5346 NNS denotes plates
T12760 5347-5351 VBD denotes were
T12758 5352-5360 VBN denotes observed
T12762 5361-5362 CD denotes 5
T12763 5363-5367 NNS denotes days
T12764 5368-5373 RB denotes later
T12765 5373-5375 , denotes ,
T12766 5375-5378 DT denotes the
T12767 5379-5384 NNS denotes cells
T12768 5385-5388 VBD denotes had
T12761 5389-5397 VBN denotes detached
T12769 5397-5398 . denotes .
T12770 5398-5528 sentence denotes Altogether, the results suggest that RMCE can give rise to p53GFP-expressing MEFs, but they die rapidly owing to p53GFP toxicity.
T12771 5399-5409 RB denotes Altogether
T12773 5409-5411 , denotes ,
T12774 5411-5414 DT denotes the
T12775 5415-5422 NNS denotes results
T12772 5423-5430 VBP denotes suggest
T12776 5431-5435 IN denotes that
T12778 5436-5440 NN denotes RMCE
T12779 5441-5444 MD denotes can
T12777 5445-5449 VB denotes give
T12780 5450-5454 NN denotes rise
T12781 5455-5457 IN denotes to
T12782 5458-5464 NN denotes p53GFP
T12784 5464-5465 HYPH denotes -
T12783 5465-5475 VBG denotes expressing
T12785 5476-5480 NNS denotes MEFs
T12786 5480-5482 , denotes ,
T12787 5482-5485 CC denotes but
T12788 5486-5490 PRP denotes they
T12789 5491-5494 VBP denotes die
T12790 5495-5502 RB denotes rapidly
T12791 5503-5508 VBG denotes owing
T12792 5509-5511 IN denotes to
T12793 5512-5518 NN denotes p53GFP
T12794 5519-5527 NN denotes toxicity
T12795 5527-5528 . denotes .
T12796 5528-5598 sentence denotes We also performed RMCE in p53RMCE/− MEFs with the p53ΔPGFP construct.
T12797 5529-5531 PRP denotes We
T12799 5532-5536 RB denotes also
T12798 5537-5546 VBD denotes performed
T12800 5547-5551 NN denotes RMCE
T12801 5552-5554 IN denotes in
T12802 5555-5562 NN denotes p53RMCE
T12804 5562-5563 HYPH denotes /
T12805 5563-5564 SYM denotes −
T12803 5565-5569 NNS denotes MEFs
T12806 5570-5574 IN denotes with
T12807 5575-5578 DT denotes the
T12809 5579-5587 NN denotes p53ΔPGFP
T12808 5588-5597 NN denotes construct
T12810 5597-5598 . denotes .
T12811 5598-5791 sentence denotes p53ΔPGFP-expressing MEFs were viable, recovered with an efficiency of ∼40%, and, as expected from ES cell experiments, expressed a p53ΔPGFP protein at much lower levels than p53WT (Figure 5B).
T12812 5599-5607 NN denotes p53ΔPGFP
T12814 5607-5608 HYPH denotes -
T12813 5608-5618 VBG denotes expressing
T12815 5619-5623 NNS denotes MEFs
T12816 5624-5628 VBD denotes were
T12817 5629-5635 JJ denotes viable
T12818 5635-5637 , denotes ,
T12819 5637-5646 VBD denotes recovered
T12820 5647-5651 IN denotes with
T12821 5652-5654 DT denotes an
T12822 5655-5665 NN denotes efficiency
T12823 5666-5668 IN denotes of
T12824 5669-5670 SYM denotes ∼
T12825 5670-5672 CD denotes 40
T12826 5672-5673 NN denotes %
T12827 5673-5675 , denotes ,
T12828 5675-5678 CC denotes and
T12829 5678-5680 , denotes ,
T12830 5680-5682 IN denotes as
T12831 5683-5691 VBN denotes expected
T12833 5692-5696 IN denotes from
T12834 5697-5699 NN denotes ES
T12835 5700-5704 NN denotes cell
T12836 5705-5716 NNS denotes experiments
T12837 5716-5718 , denotes ,
T12832 5718-5727 VBD denotes expressed
T12838 5728-5729 DT denotes a
T12840 5730-5738 NN denotes p53ΔPGFP
T12839 5739-5746 NN denotes protein
T12841 5747-5749 IN denotes at
T12842 5750-5754 RB denotes much
T12843 5755-5760 JJR denotes lower
T12844 5761-5767 NNS denotes levels
T12845 5768-5772 IN denotes than
T12846 5773-5778 NN denotes p53WT
T12847 5779-5780 -LRB- denotes (
T12849 5780-5786 NN denotes Figure
T12848 5787-5789 NN denotes 5B
T12850 5789-5790 -RRB- denotes )
T12851 5790-5791 . denotes .
T12852 5791-5873 sentence denotes Unlike WT MEFs, p53ΔP/ΔP MEFs are unable to arrest cycling after irradiation (5).
T12853 5792-5798 IN denotes Unlike
T12855 5799-5801 NN denotes WT
T12856 5802-5806 NNS denotes MEFs
T12857 5806-5808 , denotes ,
T12858 5808-5813 NN denotes p53ΔP
T12860 5813-5814 HYPH denotes /
T12859 5814-5816 NN denotes ΔP
T12861 5817-5821 NNS denotes MEFs
T12854 5822-5825 VBP denotes are
T12862 5826-5832 JJ denotes unable
T12863 5833-5835 TO denotes to
T12864 5836-5842 VB denotes arrest
T12865 5843-5850 VBG denotes cycling
T12866 5851-5856 IN denotes after
T12867 5857-5868 NN denotes irradiation
T12868 5869-5870 -LRB- denotes (
T12869 5870-5871 CD denotes 5
T12870 5871-5872 -RRB- denotes )
T12871 5872-5873 . denotes .
T12872 5873-6060 sentence denotes Likewise, we found that irradiation doses that arrested p53RMCE/− MEFs (which express a wild-type p53 from the RMCE-ready locus, see Figure 2) did not arrest p53ΔPGFP/− MEFs (Figure 5C).
T12873 5874-5882 RB denotes Likewise
T12875 5882-5884 , denotes ,
T12876 5884-5886 PRP denotes we
T12874 5887-5892 VBD denotes found
T12877 5893-5897 IN denotes that
T12879 5898-5909 NN denotes irradiation
T12880 5910-5915 NNS denotes doses
T12881 5916-5920 WDT denotes that
T12882 5921-5929 VBD denotes arrested
T12883 5930-5937 NN denotes p53RMCE
T12885 5937-5938 HYPH denotes /
T12886 5938-5939 SYM denotes −
T12884 5940-5944 NNS denotes MEFs
T12887 5945-5946 -LRB- denotes (
T12888 5946-5951 WDT denotes which
T12889 5952-5959 VBP denotes express
T12890 5960-5961 DT denotes a
T12892 5962-5966 JJ denotes wild
T12894 5966-5967 HYPH denotes -
T12893 5967-5971 NN denotes type
T12891 5972-5975 NN denotes p53
T12895 5976-5980 IN denotes from
T12896 5981-5984 DT denotes the
T12898 5985-5989 NN denotes RMCE
T12900 5989-5990 HYPH denotes -
T12899 5990-5995 JJ denotes ready
T12897 5996-6001 NN denotes locus
T12901 6001-6003 , denotes ,
T12902 6003-6006 VBP denotes see
T12903 6007-6013 NN denotes Figure
T12904 6014-6015 CD denotes 2
T12905 6015-6016 -RRB- denotes )
T12906 6017-6020 VBD denotes did
T12907 6021-6024 RB denotes not
T12878 6025-6031 VB denotes arrest
T12908 6032-6040 NN denotes p53ΔPGFP
T12910 6040-6041 HYPH denotes /
T12911 6041-6042 SYM denotes −
T12909 6043-6047 NNS denotes MEFs
T12912 6048-6049 -LRB- denotes (
T12914 6049-6055 NN denotes Figure
T12913 6056-6058 NN denotes 5C
T12915 6058-6059 -RRB- denotes )
T12916 6059-6060 . denotes .
T12917 6060-6174 sentence denotes These data show that a single RMCE-ASAP reaction in heterozygous MEFs enables detection of a recessive phenotype.
T12918 6061-6066 DT denotes These
T12919 6067-6071 NNS denotes data
T12920 6072-6076 VBP denotes show
T12921 6077-6081 IN denotes that
T12923 6082-6083 DT denotes a
T12925 6084-6090 JJ denotes single
T12926 6091-6095 NN denotes RMCE
T12928 6095-6096 HYPH denotes -
T12927 6096-6100 NN denotes ASAP
T12924 6101-6109 NN denotes reaction
T12929 6110-6112 IN denotes in
T12930 6113-6125 JJ denotes heterozygous
T12931 6126-6130 NNS denotes MEFs
T12922 6131-6138 VBZ denotes enables
T12932 6139-6148 NN denotes detection
T12933 6149-6151 IN denotes of
T12934 6152-6153 DT denotes a
T12936 6154-6163 JJ denotes recessive
T12935 6164-6173 NN denotes phenotype
T12937 6173-6174 . denotes .
T12938 6174-6377 sentence denotes The results confirm that deleting the proline rich domain leads to less active p53 with impaired cell cycle control, and also indicate that a GFP C-terminal fusion can dramatically alter p53 regulation.
T12939 6175-6178 DT denotes The
T12940 6179-6186 NNS denotes results
T12941 6187-6194 VBP denotes confirm
T12942 6195-6199 IN denotes that
T12944 6200-6208 VBG denotes deleting
T12945 6209-6212 DT denotes the
T12947 6213-6220 NN denotes proline
T12948 6221-6225 JJ denotes rich
T12946 6226-6232 NN denotes domain
T12943 6233-6238 VBZ denotes leads
T12949 6239-6241 IN denotes to
T12950 6242-6246 RBR denotes less
T12951 6247-6253 JJ denotes active
T12952 6254-6257 NN denotes p53
T12953 6258-6262 IN denotes with
T12954 6263-6271 JJ denotes impaired
T12956 6272-6276 NN denotes cell
T12957 6277-6282 NN denotes cycle
T12955 6283-6290 NN denotes control
T12958 6290-6292 , denotes ,
T12959 6292-6295 CC denotes and
T12960 6296-6300 RB denotes also
T12961 6301-6309 VBP denotes indicate
T12962 6310-6314 IN denotes that
T12964 6315-6316 DT denotes a
T12966 6317-6320 NN denotes GFP
T12967 6321-6322 NN denotes C
T12968 6322-6323 HYPH denotes -
T12969 6323-6331 JJ denotes terminal
T12965 6332-6338 NN denotes fusion
T12970 6339-6342 MD denotes can
T12971 6343-6355 RB denotes dramatically
T12963 6356-6361 VB denotes alter
T12972 6362-6365 NN denotes p53
T12973 6366-6376 NN denotes regulation
T12974 6376-6377 . denotes .
T12975 6377-6427 sentence denotes A summary of our results is presented in Table 1.
T12976 6378-6379 DT denotes A
T12977 6380-6387 NN denotes summary
T12979 6388-6390 IN denotes of
T12980 6391-6394 PRP$ denotes our
T12981 6395-6402 NNS denotes results
T12982 6403-6405 VBZ denotes is
T12978 6406-6415 VBN denotes presented
T12983 6416-6418 IN denotes in
T12984 6419-6424 NN denotes Table
T12985 6425-6426 CD denotes 1
T12986 6426-6427 . denotes .
T12987 6427-6641 sentence denotes These data report the development and implementation of an improved RMCE approach that enables efficient allele modification in ES cells to generate mice and in heterozygous MEFs to accelerate phenotypic analyses.
T12988 6428-6433 DT denotes These
T12989 6434-6438 NNS denotes data
T12990 6439-6445 VBP denotes report
T12991 6446-6449 DT denotes the
T12992 6450-6461 NN denotes development
T12993 6462-6465 CC denotes and
T12994 6466-6480 NN denotes implementation
T12995 6481-6483 IN denotes of
T12996 6484-6486 DT denotes an
T12998 6487-6495 VBN denotes improved
T12999 6496-6500 NN denotes RMCE
T12997 6501-6509 NN denotes approach
T13000 6510-6514 WDT denotes that
T13001 6515-6522 VBZ denotes enables
T13002 6523-6532 JJ denotes efficient
T13004 6533-6539 NN denotes allele
T13003 6540-6552 NN denotes modification
T13005 6553-6555 IN denotes in
T13006 6556-6558 NN denotes ES
T13007 6559-6564 NNS denotes cells
T13008 6565-6567 TO denotes to
T13009 6568-6576 VB denotes generate
T13010 6577-6581 NNS denotes mice
T13011 6582-6585 CC denotes and
T13012 6586-6588 IN denotes in
T13014 6589-6601 JJ denotes heterozygous
T13015 6602-6606 NNS denotes MEFs
T13016 6607-6609 TO denotes to
T13013 6610-6620 VB denotes accelerate
T13017 6621-6631 JJ denotes phenotypic
T13018 6632-6640 NNS denotes analyses
T13019 6640-6641 . denotes .
T13020 6641-6904 sentence denotes The success of RMCE-ASAP relied on the integrated use of inverted heterologous loxP sites, a positive/negative selection marker that preserves the germline capacity of ES cells, and, for somatic cells, the existence of a knock-out allele of the gene of interest.
T13021 6642-6645 DT denotes The
T13022 6646-6653 NN denotes success
T13024 6654-6656 IN denotes of
T13025 6657-6661 NN denotes RMCE
T13027 6661-6662 HYPH denotes -
T13026 6662-6666 NN denotes ASAP
T13023 6667-6673 VBN denotes relied
T13028 6674-6676 IN denotes on
T13029 6677-6680 DT denotes the
T13031 6681-6691 VBN denotes integrated
T13030 6692-6695 NN denotes use
T13032 6696-6698 IN denotes of
T13033 6699-6707 VBN denotes inverted
T13035 6708-6720 JJ denotes heterologous
T13036 6721-6725 NN denotes loxP
T13034 6726-6731 NNS denotes sites
T13037 6731-6733 , denotes ,
T13038 6733-6734 DT denotes a
T13040 6735-6743 JJ denotes positive
T13042 6743-6744 HYPH denotes /
T13041 6744-6752 JJ denotes negative
T13043 6753-6762 NN denotes selection
T13039 6763-6769 NN denotes marker
T13044 6770-6774 WDT denotes that
T13045 6775-6784 VBZ denotes preserves
T13046 6785-6788 DT denotes the
T13048 6789-6797 NN denotes germline
T13047 6798-6806 NN denotes capacity
T13049 6807-6809 IN denotes of
T13050 6810-6812 NN denotes ES
T13051 6813-6818 NNS denotes cells
T13052 6818-6820 , denotes ,
T13053 6820-6823 CC denotes and
T13054 6823-6825 , denotes ,
T13055 6825-6828 IN denotes for
T13057 6829-6836 JJ denotes somatic
T13058 6837-6842 NNS denotes cells
T13059 6842-6844 , denotes ,
T13060 6844-6847 DT denotes the
T13056 6848-6857 NN denotes existence
T13061 6858-6860 IN denotes of
T13062 6861-6862 DT denotes a
T13064 6863-6868 VB denotes knock
T13065 6868-6869 HYPH denotes -
T13066 6869-6872 RP denotes out
T13063 6873-6879 NN denotes allele
T13067 6880-6882 IN denotes of
T13068 6883-6886 DT denotes the
T13069 6887-6891 NN denotes gene
T13070 6892-6894 IN denotes of
T13071 6895-6903 NN denotes interest
T13072 6903-6904 . denotes .
T13073 6904-7091 sentence denotes These characteristics should make RMCE-ASAP a robust and general technology for analysis of mammalian genes under conditions that preserve normal control mechanisms in different tissues.
T13074 6905-6910 DT denotes These
T13075 6911-6926 NNS denotes characteristics
T13077 6927-6933 MD denotes should
T13076 6934-6938 VB denotes make
T13078 6939-6943 NN denotes RMCE
T13080 6943-6944 HYPH denotes -
T13079 6944-6948 NN denotes ASAP
T13082 6949-6950 DT denotes a
T13083 6951-6957 JJ denotes robust
T13084 6958-6961 CC denotes and
T13085 6962-6969 JJ denotes general
T13081 6970-6980 NN denotes technology
T13086 6981-6984 IN denotes for
T13087 6985-6993 NN denotes analysis
T13088 6994-6996 IN denotes of
T13089 6997-7006 JJ denotes mammalian
T13090 7007-7012 NNS denotes genes
T13091 7013-7018 IN denotes under
T13092 7019-7029 NNS denotes conditions
T13093 7030-7034 WDT denotes that
T13094 7035-7043 VBP denotes preserve
T13095 7044-7050 JJ denotes normal
T13097 7051-7058 NN denotes control
T13096 7059-7069 NNS denotes mechanisms
T13098 7070-7072 IN denotes in
T13099 7073-7082 JJ denotes different
T13100 7083-7090 NNS denotes tissues
T13101 7090-7091 . denotes .
T13102 7091-7251 sentence denotes In addition, RMCE-ASAP could be used to generate fibroblastic cell lines tailored for the repeated targeting of widely studied genes (p53, c-myc, NF-KB, etc.).
T13103 7092-7094 IN denotes In
T13105 7095-7103 NN denotes addition
T13106 7103-7105 , denotes ,
T13107 7105-7109 NN denotes RMCE
T13109 7109-7110 HYPH denotes -
T13108 7110-7114 NN denotes ASAP
T13110 7115-7120 MD denotes could
T13111 7121-7123 VB denotes be
T13104 7124-7128 VBN denotes used
T13112 7129-7131 TO denotes to
T13113 7132-7140 VB denotes generate
T13114 7141-7153 JJ denotes fibroblastic
T13116 7154-7158 NN denotes cell
T13115 7159-7164 NNS denotes lines
T13117 7165-7173 VBN denotes tailored
T13118 7174-7177 IN denotes for
T13119 7178-7181 DT denotes the
T13121 7182-7190 VBN denotes repeated
T13120 7191-7200 NN denotes targeting
T13122 7201-7203 IN denotes of
T13123 7204-7210 RB denotes widely
T13124 7211-7218 VBN denotes studied
T13125 7219-7224 NNS denotes genes
T13126 7225-7226 -LRB- denotes (
T13127 7226-7229 NN denotes p53
T13128 7229-7231 , denotes ,
T13129 7231-7232 NN denotes c
T13131 7232-7233 HYPH denotes -
T13130 7233-7236 NN denotes myc
T13132 7236-7238 , denotes ,
T13133 7238-7240 NN denotes NF
T13135 7240-7241 HYPH denotes -
T13134 7241-7243 NN denotes KB
T13136 7243-7245 , denotes ,
T13137 7245-7249 FW denotes etc.
T13138 7249-7250 -RRB- denotes )
T13139 7250-7251 . denotes .
R2672 T11673 T11674 det The,rationale
R2673 T11674 T11675 nsubjpass rationale,detailed
R2674 T11676 T11674 prep for,rationale
R2675 T11677 T11678 compound RMCE,ASAP
R2676 T11678 T11676 pobj ASAP,for
R2677 T11679 T11678 punct -,ASAP
R2678 T11680 T11675 auxpass is,detailed
R2679 T11681 T11675 prep in,detailed
R2680 T11682 T11681 pobj Figure,in
R2681 T11683 T11682 nummod 1,Figure
R2682 T11684 T11675 punct .,detailed
R2683 T11686 T11687 det The,step
R2684 T11687 T11689 nsubj step,requires
R2685 T11688 T11687 amod first,step
R2686 T11690 T11689 xcomp generating,requires
R2687 T11691 T11692 det a,allele
R2688 T11692 T11690 dobj allele,generating
R2689 T11693 T11692 amod floxed,allele
R2690 T11694 T11690 prep in,generating
R2691 T11695 T11696 compound ES,cells
R2692 T11696 T11694 pobj cells,in
R2693 T11697 T11698 dep that,serve
R2694 T11698 T11696 relcl serve,cells
R2695 T11699 T11698 aux will,serve
R2696 T11700 T11698 prep as,serve
R2697 T11701 T11702 det the,substrate
R2698 T11702 T11700 pobj substrate,as
R2699 T11703 T11702 prep for,substrate
R2700 T11704 T11705 amod subsequent,exchanges
R2701 T11705 T11703 pobj exchanges,for
R2702 T11706 T11707 punct (,Figure
R2703 T11707 T11689 parataxis Figure,requires
R2704 T11708 T11709 npadvmod RMCE,ready
R2705 T11709 T11711 amod ready,cell
R2706 T11710 T11709 punct -,ready
R2707 T11711 T11707 dep cell,Figure
R2708 T11712 T11711 compound ES,cell
R2709 T11713 T11707 punct ", ",Figure
R2710 T11714 T11707 nummod 1,Figure
R2711 T11715 T11707 punct ),Figure
R2712 T11716 T11689 punct .,requires
R2713 T11718 T11719 det The,strategy
R2714 T11719 T11721 nsubjpass strategy,detailed
R2715 T11720 T11719 amod targeting,strategy
R2716 T11722 T11721 auxpass is,detailed
R2717 T11723 T11721 prep in,detailed
R2718 T11724 T11723 pobj Figure,in
R2719 T11725 T11724 nummod 2,Figure
R2720 T11726 T11721 punct .,detailed
R2721 T11728 T11729 det The,frequency
R2722 T11729 T11730 nsubj frequency,was
R2723 T11731 T11729 prep of,frequency
R2724 T11732 T11731 pobj targeting,of
R2725 T11733 T11734 nummod 4,%
R2726 T11734 T11730 attr %,was
R2727 T11735 T11736 punct (,Figure
R2728 T11736 T11730 parataxis Figure,was
R2729 T11737 T11738 compound 12,300
R2730 T11738 T11740 nummod 300,clones
R2731 T11739 T11738 punct /,300
R2732 T11740 T11736 dep clones,Figure
R2733 T11741 T11742 npadvmod puromycin,resistant
R2734 T11742 T11740 amod resistant,clones
R2735 T11743 T11742 punct -,resistant
R2736 T11850 T11845 punct -,furanosyl
R2737 T11851 T11845 nummod 2,furanosyl
R2738 T11852 T11845 punct -,furanosyl
R2739 T11853 T11845 nmod fluoro,furanosyl
R2740 T11854 T11845 punct -,furanosyl
R2741 T11855 T11845 nummod 1,furanosyl
R2742 T11856 T11845 punct -,furanosyl
R2743 T11857 T11845 compound b,furanosyl
R2744 T11858 T11845 punct -,furanosyl
R2745 T11859 T11845 compound d,furanosyl
R2746 T11860 T11845 punct -,furanosyl
R2747 T11861 T11845 compound arabino,furanosyl
R2748 T11862 T11845 punct -,furanosyl
R2749 T11863 T11845 punct ),furanosyl
R2750 T11864 T11842 punct -,5
R2751 T11744 T11740 punct ", ",clones
R2752 T11865 T11840 pobj iodouracil,to
R2753 T11745 T11740 acl analyzed,clones
R2754 T11746 T11745 prep by,analyzed
R2755 T11747 T11748 compound Southern,blot
R2756 T11866 T11865 punct -,iodouracil
R2757 T11748 T11746 pobj blot,by
R2758 T11749 T11748 cc and,blot
R2759 T11750 T11751 amod long,range
R2760 T11867 T11865 punct (,iodouracil
R2761 T11751 T11753 compound range,PCR
R2762 T11752 T11751 punct -,range
R2763 T11753 T11748 conj PCR,blot
R2764 T11868 T11865 appos FIAU,iodouracil
R2765 T11754 T11736 punct ", ",Figure
R2766 T11755 T11736 nummod 2,Figure
R2767 T11756 T11736 punct ),Figure
R2768 T11869 T11839 punct ),resistant
R2769 T11757 T11730 punct .,was
R2770 T11759 T11760 nsubj We,tested
R2771 T11761 T11760 advmod next,tested
R2772 T11762 T11763 det the,efficiency
R2773 T11870 T11839 prep due,resistant
R2774 T11763 T11760 dobj efficiency,tested
R2775 T11764 T11763 prep of,efficiency
R2776 T11765 T11764 pobj RMCE,of
R2777 T11871 T11870 pcomp to,due
R2778 T11766 T11763 prep in,efficiency
R2779 T11767 T11768 compound ES,cells
R2780 T11768 T11766 pobj cells,in
R2781 T11872 T11873 compound TK,loss
R2782 T11769 T11760 punct ", ",tested
R2783 T11873 T11870 pobj loss,due
R2784 T11874 T11834 cc and,picked
R2785 T11770 T11760 advcl using,tested
R2786 T11771 T11772 det a,construct
R2787 T11772 T11770 dobj construct,using
R2788 T11875 T11834 conj analyzed,picked
R2789 T11773 T11772 compound replacement,construct
R2790 T11774 T11772 acl encoding,construct
R2791 T11876 T11877 poss their,DNA
R2792 T11775 T11774 dobj p53,encoding
R2793 T11877 T11875 dobj DNA,analyzed
R2794 T11776 T11772 acl fused,construct
R2795 T11777 T11776 prep to,fused
R2796 T11778 T11777 pobj GFP,to
R2797 T11878 T11875 prep by,analyzed
R2798 T11779 T11778 punct (,GFP
R2799 T11780 T11778 appos p53GFP,GFP
R2800 T11781 T11770 punct ),using
R2801 T11879 T11878 pobj PCR,by
R2802 T11782 T11783 aux to,enable
R2803 T11783 T11770 advcl enable,using
R2804 T11784 T11785 amod tracking,p53
R2805 T11880 T11879 cc and,PCR
R2806 T11785 T11783 dobj p53,enable
R2807 T11786 T11783 prep in,enable
R2808 T11787 T11788 amod individual,cells
R2809 T11881 T11882 compound Southern,blot
R2810 T11788 T11786 pobj cells,in
R2811 T11789 T11788 amod live,cells
R2812 T11790 T11760 punct .,tested
R2813 T11882 T11879 conj blot,PCR
R2814 T11792 T11793 advmod Importantly,used
R2815 T11883 T11834 punct .,picked
R2816 T11794 T11793 advmod however,used
R2817 T11795 T11793 punct ", ",used
R2818 T11796 T11797 compound GFP,fluorescence
R2819 T11885 T11886 advmod Strikingly,identified
R2820 T11797 T11793 nsubjpass fluorescence,used
R2821 T11798 T11793 auxpass was,used
R2822 T11799 T11793 neg not,used
R2823 T11800 T11801 aux to,screen
R2824 T11801 T11793 advcl screen,used
R2825 T11802 T11801 dobj cells,screen
R2826 T11803 T11802 prep with,cells
R2827 T11887 T11886 punct ", ",identified
R2828 T11804 T11805 amod targeted,events
R2829 T11805 T11803 pobj events,with
R2830 T11806 T11793 punct ", ",used
R2831 T11807 T11808 mark as,wanted
R2832 T11888 T11889 nummod 54,recombinants
R2833 T11808 T11793 advcl wanted,used
R2834 T11809 T11808 nsubj we,wanted
R2835 T11810 T11811 aux to,develop
R2836 T11811 T11808 xcomp develop,wanted
R2837 T11889 T11886 nsubjpass recombinants,identified
R2838 T11812 T11813 det a,method
R2839 T11813 T11811 dobj method,develop
R2840 T11814 T11813 amod general,method
R2841 T11890 T11889 amod proper,recombinants
R2842 T11815 T11816 aux to,isolate
R2843 T11816 T11811 advcl isolate,develop
R2844 T11891 T11886 auxpass were,identified
R2845 T11817 T11818 npadvmod marker,free
R2846 T11818 T11820 amod free,recombinants
R2847 T11819 T11818 punct -,free
R2848 T11892 T11886 punct ", ",identified
R2849 T11820 T11816 dobj recombinants,isolate
R2850 T11821 T11793 punct .,used
R2851 T11893 T11886 advcl indicating,identified
R2852 T11823 T11824 det The,strategy
R2853 T11824 T11826 nsubjpass strategy,detailed
R2854 T11825 T11824 compound exchange,strategy
R2855 T11894 T11895 advmod very,high
R2856 T11827 T11826 auxpass is,detailed
R2857 T11895 T11896 amod high,efficiency
R2858 T11828 T11826 prep in,detailed
R2859 T11829 T11830 compound Figure,3A
R2860 T11830 T11828 pobj 3A,in
R2861 T11896 T11893 dobj efficiency,indicating
R2862 T11831 T11826 punct .,detailed
R2863 T11833 T11834 nsubj We,picked
R2864 T11897 T11896 compound RMCE,efficiency
R2865 T11835 T11836 nummod 65,clones
R2866 T11836 T11834 dobj clones,picked
R2867 T11898 T11899 punct (,%
R2868 T11837 T11838 compound ES,cell
R2869 T11838 T11836 compound cell,clones
R2870 T11839 T11836 relcl resistant,clones
R2871 T11899 T11896 parataxis %,efficiency
R2872 T11840 T11839 prep to,resistant
R2873 T11841 T11842 nummod 1,5
R2874 T11900 T11899 nummod 83,%
R2875 T11842 T11865 nummod 5,iodouracil
R2876 T11843 T11842 punct -,5
R2877 T11901 T11899 punct ),%
R2878 T11844 T11845 punct (,furanosyl
R2879 T11845 T11842 parataxis furanosyl,5
R2880 T11846 T11845 punct -,furanosyl
R2881 T11902 T11886 punct .,identified
R2882 T11847 T11845 nummod 2,furanosyl
R2883 T11848 T11845 punct -,furanosyl
R2884 T11849 T11845 nmod deoxy,furanosyl
R2885 T11904 T11905 nsubj RMCE,proved
R2886 T11906 T11905 advmod also,proved
R2887 T11907 T11908 aux to,be
R2888 T11908 T11905 xcomp be,proved
R2889 T11909 T11908 acomp precise,be
R2890 T11910 T11908 punct ", ",be
R2891 T11955 T11954 amod expected,size
R2892 T11956 T11954 punct (,size
R2893 T11911 T11912 mark as,detected
R2894 T11957 T11958 quantmod ca.,80
R2895 T11958 T11959 nummod 80,kDa
R2896 T11912 T11908 advcl detected,be
R2897 T11959 T11954 appos kDa,size
R2898 T11960 T11934 punct ),analyzed
R2899 T11961 T11934 punct .,analyzed
R2900 T11913 T11914 det no,bands
R2901 T11963 T11964 advmod Surprisingly,altered
R2902 T11914 T11912 nsubjpass bands,detected
R2903 T11964 T11973 ccomp altered,were
R2904 T11965 T11964 punct ", ",altered
R2905 T11966 T11967 det the,fusion
R2906 T11915 T11914 amod aberrant,bands
R2907 T11967 T11964 nsubj fusion,altered
R2908 T11968 T11967 prep of,fusion
R2909 T11916 T11912 auxpass were,detected
R2910 T11969 T11968 pobj GFP,of
R2911 T11970 T11967 prep to,fusion
R2912 T11971 T11970 pobj p53,to
R2913 T11917 T11912 prep in,detected
R2914 T11972 T11964 advmod apparently,altered
R2915 T11974 T11975 compound p53,stability
R2916 T11918 T11917 pobj PCR,in
R2917 T11975 T11964 dobj stability,altered
R2918 T11976 T11973 punct : ,were
R2919 T11977 T11978 amod steady,state
R2920 T11919 T11918 cc and,PCR
R2921 T11978 T11980 compound state,levels
R2922 T11979 T11978 punct -,state
R2923 T11980 T11973 nsubj levels,were
R2924 T11920 T11921 compound Southern,blots
R2925 T11981 T11980 prep of,levels
R2926 T11982 T11981 pobj p53GFP,of
R2927 T11983 T11984 advmod much,higher
R2928 T11984 T11973 acomp higher,were
R2929 T11985 T11984 prep than,higher
R2930 T11986 T11985 pobj those,than
R2931 T11987 T11986 prep of,those
R2932 T11921 T11918 conj blots,PCR
R2933 T11988 T11989 amod wild,type
R2934 T11989 T11991 compound type,p53
R2935 T11990 T11989 punct -,type
R2936 T11922 T11923 punct (,3A
R2937 T11991 T11987 pobj p53,of
R2938 T11992 T11991 punct (,p53
R2939 T11923 T11912 parataxis 3A,detected
R2940 T11993 T11991 appos p53WT,p53
R2941 T11994 T11991 punct ),p53
R2942 T11995 T11991 prep in,p53
R2943 T11924 T11923 compound Figure,3A
R2944 T11996 T11997 amod unstressed,cells
R2945 T11997 T11995 pobj cells,in
R2946 T11998 T11973 punct ", ",were
R2947 T11925 T11923 punct ),3A
R2948 T11999 T11973 cc and,were
R2949 T12000 T12001 aux did,vary
R2950 T12001 T11973 conj vary,were
R2951 T11926 T11905 punct .,proved
R2952 T12002 T12001 neg not,vary
R2953 T12003 T12001 advmod significantly,vary
R2954 T11928 T11929 nmod p53,GFP
R2955 T12004 T12001 prep after,vary
R2956 T12005 T12006 compound DNA,damage
R2957 T12006 T12004 pobj damage,after
R2958 T12007 T12001 punct ", ",vary
R2959 T12008 T12009 mark so,were
R2960 T11929 T11932 compound GFP,clones
R2961 T12009 T12001 advcl were,vary
R2962 T12010 T12009 mark that,were
R2963 T12011 T12012 det the,levels
R2964 T11930 T11929 punct +,GFP
R2965 T12012 T12009 nsubj levels,were
R2966 T12013 T12012 prep for,levels
R2967 T12014 T12015 preconj both,p53WT
R2968 T12015 T12013 pobj p53WT,for
R2969 T11931 T11929 punct /,GFP
R2970 T12016 T12015 cc and,p53WT
R2971 T12017 T12015 conj p53GFP,p53WT
R2972 T12018 T12009 acomp similar,were
R2973 T11932 T11934 nsubjpass clones,analyzed
R2974 T12019 T12009 prep after,were
R2975 T12020 T12021 compound adriamycin,treatment
R2976 T12021 T12019 pobj treatment,after
R2977 T11933 T11932 compound ES,clones
R2978 T12022 T12023 punct (,3A
R2979 T12023 T12009 parataxis 3A,were
R2980 T12024 T12023 compound Figure,3A
R2981 T11935 T11934 auxpass were,analyzed
R2982 T12025 T12023 punct ),3A
R2983 T12026 T11973 punct .,were
R2984 T12028 T12029 nummod Six,clones
R2985 T11936 T11934 prep by,analyzed
R2986 T12029 T12035 nsubjpass clones,injected
R2987 T12030 T12029 amod independent,clones
R2988 T11937 T11938 compound western,blot
R2989 T12031 T12032 nmod p53,GFP
R2990 T12032 T12029 compound GFP,clones
R2991 T12033 T12032 punct +,GFP
R2992 T11938 T11936 pobj blot,by
R2993 T12034 T12032 punct /,GFP
R2994 T12036 T12035 auxpass were,injected
R2995 T11939 T11934 prep with,analyzed
R2996 T12037 T12035 prep into,injected
R2997 T12038 T12037 pobj blastocysts,into
R2998 T12039 T12035 cc and,injected
R2999 T11940 T11941 det an,antibody
R3000 T12040 T12035 conj transferred,injected
R3001 T12041 T12040 prep to,transferred
R3002 T12042 T12043 amod pseudo,pregnant
R3003 T11941 T11939 pobj antibody,with
R3004 T12043 T12045 amod pregnant,females
R3005 T12044 T12043 punct -,pregnant
R3006 T11942 T11941 prep against,antibody
R3007 T12045 T12041 pobj females,to
R3008 T12046 T12040 advcl using,transferred
R3009 T12047 T12048 amod standard,procedures
R3010 T11943 T11942 pobj p53,against
R3011 T12048 T12046 dobj procedures,using
R3012 T12049 T12035 punct .,injected
R3013 T11944 T11934 punct ", ",analyzed
R3014 T12051 T12052 advmod Strikingly,obtained
R3015 T12053 T12052 punct ", ",obtained
R3016 T11945 T11934 cc and,analyzed
R3017 T12054 T12055 det no,pregnancies
R3018 T12055 T12052 nsubjpass pregnancies,obtained
R3019 T12056 T12052 auxpass were,obtained
R3020 T11946 T11934 conj found,analyzed
R3021 T12057 T12052 punct .,obtained
R3022 T12059 T12060 nsubjpass It,shown
R3023 T11947 T11948 aux to,express
R3024 T12060 T12063 ccomp shown,contain
R3025 T11948 T11946 xcomp express,found
R3026 T11949 T11950 det an,band
R3027 T12061 T12060 aux has,shown
R3028 T11950 T11948 dobj band,express
R3029 T12062 T12060 auxpass been,shown
R3030 T12064 T12065 mark that,regulated
R3031 T12065 T12060 ccomp regulated,shown
R3032 T12066 T12067 det the,pathway
R3033 T12067 T12065 nsubjpass pathway,regulated
R3034 T11951 T11950 amod additional,band
R3035 T12068 T12067 compound p53,pathway
R3036 T12069 T12065 auxpass is,regulated
R3037 T12070 T12071 advmod very,differently
R3038 T12071 T12065 advmod differently,regulated
R3039 T11952 T11948 prep at,express
R3040 T12072 T12065 prep in,regulated
R3041 T12073 T12074 nmod ES,cells
R3042 T12074 T12072 pobj cells,in
R3043 T11953 T11954 det the,size
R3044 T12075 T12073 cc and,ES
R3045 T12076 T12073 conj somatic,ES
R3046 T11954 T11952 pobj size,at
R3047 T12077 T12063 punct : ,contain
R3048 T12078 T12079 compound ES,cells
R3049 T12079 T12063 nsubj cells,contain
R3050 T12167 T12168 det this,possibility
R3051 T12080 T12081 advmod relatively,high
R3052 T12081 T12082 amod high,levels
R3053 T12082 T12063 dobj levels,contain
R3054 T12083 T12082 compound p53,levels
R3055 T12084 T12063 cc and,contain
R3056 T12168 T12165 dobj possibility,test
R3057 T12085 T12063 conj lack,contain
R3058 T12086 T12087 det the,responses
R3059 T12087 T12085 dobj responses,lack
R3060 T12169 T12166 punct ", ",performed
R3061 T12088 T12087 nmod p53,responses
R3062 T12089 T12088 punct -,p53
R3063 T12090 T12088 amod mediated,p53
R3064 T12170 T12166 nsubj we,performed
R3065 T12091 T12092 compound DNA,damage
R3066 T12092 T12087 compound damage,responses
R3067 T12171 T12166 dobj RMCE,performed
R3068 T12093 T12087 acl found,responses
R3069 T12094 T12093 prep in,found
R3070 T12095 T12096 amod somatic,cells
R3071 T12172 T12166 prep with,performed
R3072 T12096 T12094 pobj cells,in
R3073 T12097 T12098 punct (,18
R3074 T12098 T12085 parataxis 18,lack
R3075 T12099 T12098 punct ),18
R3076 T12173 T12174 det a,gene
R3077 T12100 T12063 punct .,contain
R3078 T12174 T12172 pobj gene,with
R3079 T12102 T12103 nsubj This,suggested
R3080 T12175 T12176 compound p53,fusion
R3081 T12103 T12120 ccomp suggested,speculate
R3082 T12104 T12103 punct ", ",suggested
R3083 T12105 T12103 advmod together,suggested
R3084 T12106 T12105 prep with,together
R3085 T12107 T12108 det the,observation
R3086 T12108 T12106 pobj observation,with
R3087 T12176 T12174 compound fusion,gene
R3088 T12109 T12110 mark that,decrease
R3089 T12110 T12108 acl decrease,observation
R3090 T12111 T12112 compound p53,levels
R3091 T12177 T12178 prep in,deleted
R3092 T12112 T12110 nsubj levels,decrease
R3093 T12113 T12110 prep during,decrease
R3094 T12114 T12115 compound mouse,embryogenesis
R3095 T12178 T12174 advcl deleted,gene
R3096 T12115 T12113 pobj embryogenesis,during
R3097 T12116 T12117 punct (,19
R3098 T12179 T12177 pobj which,in
R3099 T12117 T12110 parataxis 19,decrease
R3100 T12118 T12117 punct ),19
R3101 T12119 T12103 punct ", ",suggested
R3102 T12180 T12181 det the,domain
R3103 T12121 T12122 det an,explanation
R3104 T12181 T12178 nsubjpass domain,deleted
R3105 T12122 T12103 dobj explanation,suggested
R3106 T12123 T12122 prep for,explanation
R3107 T12124 T12125 det the,lack
R3108 T12182 T12181 nmod p53,domain
R3109 T12125 T12123 pobj lack,for
R3110 T12126 T12125 amod observed,lack
R3111 T12127 T12125 prep of,lack
R3112 T12183 T12184 npadvmod proline,rich
R3113 T12128 T12127 pobj pregnancies,of
R3114 T12129 T12120 punct : ,speculate
R3115 T12130 T12120 nsubj we,speculate
R3116 T12184 T12181 amod rich,domain
R3117 T12131 T12132 mark that,prevented
R3118 T12132 T12120 ccomp prevented,speculate
R3119 T12185 T12184 punct -,rich
R3120 T12133 T12134 det the,levels
R3121 T12134 T12132 nsubj levels,prevented
R3122 T12135 T12134 amod high,levels
R3123 T12186 T12181 punct (,domain
R3124 T12136 T12134 prep of,levels
R3125 T12137 T12136 pobj p53GFP,of
R3126 T12187 T12181 appos PRD,domain
R3127 T12138 T12134 prep in,levels
R3128 T12139 T12140 det the,cells
R3129 T12140 T12138 pobj cells,in
R3130 T12141 T12140 compound ES,cells
R3131 T12188 T12178 punct ),deleted
R3132 T12142 T12140 acl injected,cells
R3133 T12143 T12142 prep into,injected
R3134 T12144 T12143 pobj blastocysts,into
R3135 T12145 T12132 aux might,prevented
R3136 T12146 T12132 aux have,prevented
R3137 T12189 T12178 auxpass was,deleted
R3138 T12147 T12148 amod normal,development
R3139 T12148 T12132 dobj development,prevented
R3140 T12149 T12148 amod embryonic,development
R3141 T12190 T12191 punct (,p53ΔP
R3142 T12150 T12151 prep once,began
R3143 T12151 T12132 advcl began,prevented
R3144 T12152 T12153 det these,cells
R3145 T12191 T12178 parataxis p53ΔP,deleted
R3146 T12153 T12151 nsubj cells,began
R3147 T12154 T12155 aux to,differentiate
R3148 T12155 T12151 xcomp differentiate,began
R3149 T12192 T12191 punct ),p53ΔP
R3150 T12156 T12151 cc and,began
R3151 T12157 T12158 det the,pathway
R3152 T12193 T12178 punct ", ",deleted
R3153 T12158 T12160 nsubj pathway,became
R3154 T12159 T12158 compound p53,pathway
R3155 T12160 T12151 conj became,began
R3156 T12194 T12178 cc and,deleted
R3157 T12161 T12160 acomp functional,became
R3158 T12162 T12120 punct .,speculate
R3159 T12195 T12196 det the,p53ΔP
R3160 T12164 T12165 aux To,test
R3161 T12165 T12166 advcl test,performed
R3162 T12196 T12197 nsubjpass p53ΔP,fused
R3163 T12197 T12178 conj fused,deleted
R3164 T12198 T12197 auxpass was,fused
R3165 T12273 T12272 compound ES,cells
R3166 T12274 T12275 punct (,3B
R3167 T12199 T12197 prep to,fused
R3168 T12275 T12257 parataxis 3B,revealed
R3169 T12276 T12275 compound Figure,3B
R3170 T12277 T12275 punct ),3B
R3171 T12200 T12199 pobj GFP,to
R3172 T12278 T12257 punct .,revealed
R3173 T12201 T12166 punct .,performed
R3174 T12280 T12281 mark As,expected
R3175 T12203 T12204 nsubj We,used
R3176 T12281 T12282 advcl expected,correlated
R3177 T12282 T12287 ccomp correlated,was
R3178 T12283 T12282 punct ", ",correlated
R3179 T12205 T12206 det this,mutant
R3180 T12284 T12285 det the,deletion
R3181 T12285 T12282 nsubj deletion,correlated
R3182 T12286 T12285 compound PRD,deletion
R3183 T12206 T12204 dobj mutant,used
R3184 T12288 T12282 prep with,correlated
R3185 T12289 T12290 amod lower,levels
R3186 T12290 T12288 pobj levels,with
R3187 T12291 T12290 compound expression,levels
R3188 T12292 T12287 punct : ,was
R3189 T12207 T12208 mark because,decreases
R3190 T12293 T12287 nsubj p53ΔPGFP,was
R3191 T12294 T12295 advmod much,less
R3192 T12295 T12296 advmod less,abundant
R3193 T12208 T12204 advcl decreases,used
R3194 T12296 T12287 acomp abundant,was
R3195 T12297 T12296 prep than,abundant
R3196 T12298 T12297 pobj p53WT,than
R3197 T12209 T12208 csubj deleting,decreases
R3198 T12299 T12298 prep in,p53WT
R3199 T12300 T12301 det all,clones
R3200 T12301 T12299 pobj clones,in
R3201 T12210 T12211 det the,domain
R3202 T12302 T12303 nmod p53,ΔPGFP
R3203 T12303 T12301 compound ΔPGFP,clones
R3204 T12304 T12303 punct +,ΔPGFP
R3205 T12211 T12209 dobj domain,deleting
R3206 T12305 T12303 punct /,ΔPGFP
R3207 T12306 T12307 punct (,3B
R3208 T12307 T12287 parataxis 3B,was
R3209 T12212 T12213 npadvmod proline,rich
R3210 T12308 T12307 compound Figure,3B
R3211 T12309 T12307 punct ),3B
R3212 T12310 T12287 punct .,was
R3213 T12213 T12211 amod rich,domain
R3214 T12312 T12313 nsubj We,determined
R3215 T12214 T12213 punct -,rich
R3216 T12314 T12313 advmod next,determined
R3217 T12315 T12316 mark whether,generate
R3218 T12215 T12208 dobj stability,decreases
R3219 T12316 T12313 ccomp generate,determined
R3220 T12317 T12318 nmod p53,ΔPGFP
R3221 T12216 T12208 cc and,decreases
R3222 T12318 T12321 compound ΔPGFP,cells
R3223 T12217 T12208 conj compromises,decreases
R3224 T12319 T12318 punct +,ΔPGFP
R3225 T12218 T12219 compound DNA,damage
R3226 T12320 T12318 punct /,ΔPGFP
R3227 T12321 T12316 nsubj cells,generate
R3228 T12219 T12221 compound damage,responses
R3229 T12322 T12321 compound ES,cells
R3230 T12220 T12219 punct -,damage
R3231 T12323 T12316 aux could,generate
R3232 T12324 T12325 amod chimeric,mice
R3233 T12325 T12316 dobj mice,generate
R3234 T12326 T12316 cc and,generate
R3235 T12221 T12217 dobj responses,compromises
R3236 T12327 T12316 conj transmit,generate
R3237 T12328 T12329 det the,allele
R3238 T12329 T12327 dobj allele,transmit
R3239 T12330 T12329 amod modified,allele
R3240 T12331 T12327 prep through,transmit
R3241 T12332 T12333 det the,germline
R3242 T12222 T12223 advmod in,vivo
R3243 T12333 T12331 pobj germline,through
R3244 T12334 T12313 punct .,determined
R3245 T12223 T12217 advmod vivo,compromises
R3246 T12336 T12337 nummod Two,clones
R3247 T12337 T12344 nsubjpass clones,injected
R3248 T12338 T12339 nmod p53,ΔPGFP
R3249 T12224 T12225 punct (,5
R3250 T12339 T12337 compound ΔPGFP,clones
R3251 T12340 T12339 punct +,ΔPGFP
R3252 T12341 T12339 punct /,ΔPGFP
R3253 T12225 T12217 parataxis 5,compromises
R3254 T12342 T12343 compound ES,cell
R3255 T12343 T12337 compound cell,clones
R3256 T12226 T12225 punct ),5
R3257 T12345 T12344 auxpass were,injected
R3258 T12346 T12344 prep into,injected
R3259 T12227 T12204 punct .,used
R3260 T12347 T12346 pobj blastocysts,into
R3261 T12348 T12344 cc and,injected
R3262 T12349 T12350 advmod highly,chimeric
R3263 T12229 T12230 prep According,prevent
R3264 T12350 T12351 amod chimeric,%
R3265 T12351 T12355 amod %,mice
R3266 T12352 T12351 punct (,%
R3267 T12353 T12351 punct >,%
R3268 T12231 T12229 prep to,According
R3269 T12354 T12351 nummod 80,%
R3270 T12355 T12357 nsubjpass mice,obtained
R3271 T12356 T12351 punct ),%
R3272 T12357 T12344 conj obtained,injected
R3273 T12232 T12233 poss our,hypothesis
R3274 T12358 T12357 auxpass were,obtained
R3275 T12359 T12357 punct .,obtained
R3276 T12233 T12231 pobj hypothesis,to
R3277 T12361 T12362 compound Heterozygote,pups
R3278 T12362 T12363 nsubjpass pups,recovered
R3279 T12234 T12230 punct ", ",prevent
R3280 T12364 T12363 auxpass were,recovered
R3281 T12365 T12363 prep from,recovered
R3282 T12235 T12236 det this,mutant
R3283 T12366 T12367 det the,mating
R3284 T12367 T12365 pobj mating,from
R3285 T12236 T12230 nsubj mutant,prevent
R3286 T12237 T12236 amod hypomorphic,mutant
R3287 T12368 T12369 det the,chimeras
R3288 T12369 T12367 nmod chimeras,mating
R3289 T12370 T12367 prep with,mating
R3290 T12371 T12372 compound WT,mice
R3291 T12372 T12370 pobj mice,with
R3292 T12373 T12374 punct (,3C
R3293 T12374 T12363 parataxis 3C,recovered
R3294 T12238 T12230 aux should,prevent
R3295 T12375 T12374 compound Figure,3C
R3296 T12376 T12374 punct ),3C
R3297 T12377 T12363 punct .,recovered
R3298 T12239 T12230 neg not,prevent
R3299 T12240 T12241 amod embryonic,development
R3300 T12241 T12230 dobj development,prevent
R3301 T12242 T12230 punct .,prevent
R3302 T12379 T12380 det These,data
R3303 T12380 T12381 nsubj data,demonstrate
R3304 T12244 T12245 nsubj RMCE,was
R3305 T12382 T12383 mark that,is
R3306 T12383 T12381 ccomp is,demonstrate
R3307 T12384 T12385 npadvmod marker,free
R3308 T12246 T12244 prep with,RMCE
R3309 T12385 T12387 amod free,RMCE
R3310 T12386 T12385 punct -,free
R3311 T12387 T12383 nsubj RMCE,is
R3312 T12388 T12389 advmod very,efficient
R3313 T12247 T12248 det a,plasmid
R3314 T12389 T12383 acomp efficient,is
R3315 T12390 T12383 prep in,is
R3316 T12391 T12392 compound ES,cells
R3317 T12248 T12246 pobj plasmid,with
R3318 T12392 T12390 pobj cells,in
R3319 T12393 T12383 cc and,is
R3320 T12394 T12383 conj allows,is
R3321 T12249 T12248 compound p53ΔPGFP,plasmid
R3322 T12395 T12396 compound germline,transmission
R3323 T12396 T12394 dobj transmission,allows
R3324 T12397 T12396 prep of,transmission
R3325 T12250 T12248 compound replacement,plasmid
R3326 T12398 T12399 det a,mutation
R3327 T12399 T12397 pobj mutation,of
R3328 T12251 T12245 advmod again,was
R3329 T12400 T12399 amod targeted,mutation
R3330 T12401 T12402 punct (,see
R3331 T12402 T12394 parataxis see,allows
R3332 T12252 T12253 advmod very,efficient
R3333 T12403 T12402 dobj Figure,see
R3334 T12404 T12403 nummod 1,Figure
R3335 T12405 T12403 punct ", ",Figure
R3336 T12253 T12245 acomp efficient,was
R3337 T12406 T12407 compound path,A
R3338 T12407 T12403 appos A,Figure
R3339 T12254 T12245 cc and,was
R3340 T12408 T12402 punct ),see
R3341 T12409 T12381 punct .,demonstrate
R3342 T12411 T12412 nsubj We,determined
R3343 T12255 T12256 compound western,blots
R3344 T12413 T12412 advmod next,determined
R3345 T12414 T12415 mark whether,used
R3346 T12415 T12412 ccomp used,determined
R3347 T12256 T12257 nsubj blots,revealed
R3348 T12416 T12417 det the,approach
R3349 T12417 T12415 nsubjpass approach,used
R3350 T12418 T12417 compound RMCE,approach
R3351 T12257 T12245 conj revealed,was
R3352 T12419 T12415 aux could,used
R3353 T12420 T12415 auxpass be,used
R3354 T12421 T12422 aux to,target
R3355 T12258 T12259 det an,band
R3356 T12422 T12415 advcl target,used
R3357 T12423 T12422 dobj mutations,target
R3358 T12424 T12423 prep at,mutations
R3359 T12259 T12257 dobj band,revealed
R3360 T12425 T12426 det the,allele
R3361 T12426 T12424 pobj allele,at
R3362 T12427 T12426 compound p53,allele
R3363 T12260 T12259 amod additional,band
R3364 T12428 T12423 prep in,mutations
R3365 T12429 T12430 amod somatic,cells
R3366 T12430 T12428 pobj cells,in
R3367 T12261 T12259 prep of,band
R3368 T12431 T12432 punct (,Figure
R3369 T12432 T12415 parataxis Figure,used
R3370 T12433 T12432 nummod 1,Figure
R3371 T12262 T12263 det the,weight
R3372 T12434 T12432 punct ", ",Figure
R3373 T12435 T12436 compound path,B
R3374 T12436 T12432 appos B,Figure
R3375 T12263 T12261 pobj weight,of
R3376 T12437 T12432 punct ),Figure
R3377 T12438 T12412 punct .,determined
R3378 T12264 T12263 amod predicted,weight
R3379 T12440 T12441 nsubj We,verified
R3380 T12265 T12263 amod molecular,weight
R3381 T12442 T12441 advmod first,verified
R3382 T12443 T12444 mark that,transmitted
R3383 T12266 T12267 advmod only,in
R3384 T12267 T12257 prep in,revealed
R3385 T12444 T12441 ccomp transmitted,verified
R3386 T12445 T12446 det the,locus
R3387 T12268 T12269 nmod p53,ΔPGFP
R3388 T12446 T12444 nsubjpass locus,transmitted
R3389 T12447 T12448 npadvmod RMCE,ready
R3390 T12269 T12272 compound ΔPGFP,cells
R3391 T12448 T12446 amod ready,locus
R3392 T12449 T12448 punct -,ready
R3393 T12450 T12446 compound p53,locus
R3394 T12451 T12446 punct (,locus
R3395 T12452 T12446 appos p53RMCE,locus
R3396 T12453 T12444 punct ),transmitted
R3397 T12270 T12269 punct +,ΔPGFP
R3398 T12454 T12444 aux could,transmitted
R3399 T12455 T12444 auxpass be,transmitted
R3400 T12456 T12444 prep through,transmitted
R3401 T12271 T12269 punct /,ΔPGFP
R3402 T12457 T12458 det the,germline
R3403 T12458 T12456 pobj germline,through
R3404 T12459 T12444 prep by,transmitted
R3405 T12272 T12267 pobj cells,in
R3406 T12460 T12459 pcomp mating,by
R3407 T12461 T12462 nmod p53RMCE,chimeras
R3408 T12483 T12481 nsubj this,allowed
R3409 T12462 T12460 dobj chimeras,mating
R3410 T12463 T12461 punct /,p53RMCE
R3411 T12464 T12461 punct +,p53RMCE
R3412 T12465 T12460 prep with,mating
R3413 T12466 T12467 nmod p53,mice
R3414 T12484 T12485 nsubj us,generate
R3415 T12467 T12465 pobj mice,with
R3416 T12468 T12466 punct +,p53
R3417 T12469 T12466 punct /,p53
R3418 T12485 T12481 ccomp generate,allowed
R3419 T12470 T12466 punct −,p53
R3420 T12471 T12472 punct (,20
R3421 T12472 T12444 parataxis 20,transmitted
R3422 T12486 T12485 aux to,generate
R3423 T12473 T12472 punct ),20
R3424 T12474 T12475 punct (,Figure
R3425 T12475 T12441 parataxis Figure,verified
R3426 T12487 T12488 nmod p53RMCE,MEFs
R3427 T12476 T12475 nummod 4,Figure
R3428 T12477 T12475 punct ),Figure
R3429 T12478 T12441 punct .,verified
R3430 T12488 T12485 dobj MEFs,generate
R3431 T12480 T12481 advmod Importantly,allowed
R3432 T12489 T12487 punct /,p53RMCE
R3433 T12482 T12481 punct ", ",allowed
R3434 T12490 T12487 punct −,p53RMCE
R3435 T12491 T12488 punct ", ",MEFs
R3436 T12492 T12493 dep which,used
R3437 T12493 T12488 relcl used,MEFs
R3438 T12589 T12588 pobj MEFs,in
R3439 T12590 T12578 auxpass is,activated
R3440 T12494 T12493 auxpass were,used
R3441 T12591 T12578 neg not,activated
R3442 T12592 T12578 advmod readily,activated
R3443 T12593 T12578 prep in,activated
R3444 T12594 T12595 compound ES,cells
R3445 T12595 T12593 pobj cells,in
R3446 T12596 T12597 punct (,18
R3447 T12495 T12496 aux to,test
R3448 T12597 T12578 parataxis 18,activated
R3449 T12598 T12597 punct ),18
R3450 T12599 T12578 punct .,activated
R3451 T12496 T12493 advcl test,used
R3452 T12601 T12602 det The,results
R3453 T12497 T12496 dobj RMCE,test
R3454 T12602 T12603 nsubj results,suggest
R3455 T12604 T12602 advmod above,results
R3456 T12498 T12496 prep at,test
R3457 T12605 T12606 mark that,be
R3458 T12606 T12603 ccomp be,suggest
R3459 T12607 T12608 amod high,levels
R3460 T12499 T12500 det the,locus
R3461 T12608 T12606 nsubj levels,be
R3462 T12609 T12608 prep of,levels
R3463 T12500 T12498 pobj locus,at
R3464 T12610 T12609 pobj p53GFP,of
R3465 T12611 T12606 aux could,be
R3466 T12612 T12606 dep tolerated,be
R3467 T12501 T12500 compound p53,locus
R3468 T12613 T12612 agent by,tolerated
R3469 T12614 T12615 compound ES,cells
R3470 T12615 T12613 pobj cells,by
R3471 T12616 T12612 cc but,tolerated
R3472 T12502 T12496 prep in,test
R3473 T12617 T12612 conj toxic,tolerated
R3474 T12618 T12617 prep to,toxic
R3475 T12503 T12504 amod somatic,cells
R3476 T12619 T12618 pobj MEFs,to
R3477 T12620 T12606 punct ", ",be
R3478 T12621 T12622 mark so,failed
R3479 T12504 T12502 pobj cells,in
R3480 T12622 T12606 advcl failed,be
R3481 T12505 T12481 punct .,allowed
R3482 T12623 T12622 mark that,failed
R3483 T12624 T12622 nsubj MEFs,failed
R3484 T12507 T12508 nsubj We,attempted
R3485 T12625 T12626 prep in,occurred
R3486 T12626 T12624 advcl occurred,MEFs
R3487 T12627 T12625 pobj which,in
R3488 T12628 T12629 det an,RMCE
R3489 T12509 T12508 advmod first,attempted
R3490 T12629 T12626 nsubj RMCE,occurred
R3491 T12630 T12626 aux had,occurred
R3492 T12631 T12632 aux to,proliferate
R3493 T12632 T12622 xcomp proliferate,failed
R3494 T12510 T12508 dobj RMCE,attempted
R3495 T12633 T12603 punct .,suggest
R3496 T12635 T12636 aux To,test
R3497 T12636 T12637 advcl test,electroporated
R3498 T12511 T12508 prep in,attempted
R3499 T12638 T12639 det this,possibility
R3500 T12639 T12636 dobj possibility,test
R3501 T12640 T12637 punct ", ",electroporated
R3502 T12512 T12511 pobj MEFs,in
R3503 T12641 T12642 nmod p53RMCE,MEFs
R3504 T12642 T12637 nsubjpass MEFs,electroporated
R3505 T12643 T12641 punct /,p53RMCE
R3506 T12513 T12508 prep by,attempted
R3507 T12644 T12641 punct −,p53RMCE
R3508 T12645 T12637 auxpass were,electroporated
R3509 T12646 T12637 prep with,electroporated
R3510 T12514 T12513 pcomp electroporating,by
R3511 T12647 T12648 det the,construct
R3512 T12648 T12646 pobj construct,with
R3513 T12649 T12648 compound p53GFP,construct
R3514 T12515 T12516 nmod p53RMCE,MEFs
R3515 T12650 T12648 compound replacement,construct
R3516 T12651 T12637 prep with,electroporated
R3517 T12516 T12514 dobj MEFs,electroporating
R3518 T12652 T12651 cc or,with
R3519 T12653 T12651 conj without,with
R3520 T12654 T12655 det a,plasmid
R3521 T12517 T12515 punct /,p53RMCE
R3522 T12655 T12653 pobj plasmid,without
R3523 T12656 T12657 compound Cre,expression
R3524 T12657 T12655 compound expression,plasmid
R3525 T12518 T12515 punct −,p53RMCE
R3526 T12658 T12657 punct -,expression
R3527 T12659 T12637 punct ", ",electroporated
R3528 T12660 T12661 advmod then,analyzed
R3529 T12661 T12637 dep analyzed,electroporated
R3530 T12519 T12514 prep with,electroporating
R3531 T12662 T12661 prep by,analyzed
R3532 T12663 T12664 compound fluorescence,microscopy
R3533 T12520 T12521 det a,plasmid
R3534 T12664 T12662 pobj microscopy,by
R3535 T12665 T12666 nummod 48,h
R3536 T12666 T12667 npadvmod h,after
R3537 T12521 T12519 pobj plasmid,with
R3538 T12667 T12661 prep after,analyzed
R3539 T12668 T12667 pobj electroporation,after
R3540 T12669 T12637 punct .,electroporated
R3541 T12522 T12523 compound Cre,expression
R3542 T12671 T12672 det The,experiment
R3543 T12672 T12673 nsubjpass experiment,done
R3544 T12523 T12521 compound expression,plasmid
R3545 T12674 T12673 auxpass was,done
R3546 T12675 T12673 prep without,done
R3547 T12524 T12523 punct -,expression
R3548 T12676 T12675 pobj selection,without
R3549 T12677 T12678 aux to,enable
R3550 T12678 T12673 advcl enable,done
R3551 T12679 T12678 dobj observation,enable
R3552 T12680 T12679 prep of,observation
R3553 T12525 T12521 cc and,plasmid
R3554 T12681 T12680 pobj cells,of
R3555 T12682 T12678 prep under,enable
R3556 T12526 T12527 det the,plasmid
R3557 T12683 T12682 pobj conditions,under
R3558 T12684 T12685 advmod where,derive
R3559 T12685 T12683 relcl derive,conditions
R3560 T12686 T12687 det a,failure
R3561 T12527 T12521 conj plasmid,plasmid
R3562 T12687 T12685 nsubj failure,derive
R3563 T12688 T12689 aux to,proliferate
R3564 T12528 T12527 compound p53GFP,plasmid
R3565 T12689 T12687 acl proliferate,failure
R3566 T12690 T12685 aux would,derive
R3567 T12691 T12685 neg not,derive
R3568 T12529 T12521 punct ", ",plasmid
R3569 T12692 T12685 prep from,derive
R3570 T12693 T12694 nmod FIAU,toxicity
R3571 T12694 T12692 pobj toxicity,from
R3572 T12530 T12521 acl followed,plasmid
R3573 T12531 T12530 prep by,followed
R3574 T12532 T12531 pobj selection,by
R3575 T12695 T12693 cc or,FIAU
R3576 T12533 T12532 prep with,selection
R3577 T12696 T12693 conj ganciclovir,FIAU
R3578 T12697 T12692 cc but,from
R3579 T12698 T12697 advmod rather,but
R3580 T12534 T12533 pobj FIAU,with
R3581 T12699 T12700 advmod solely,from
R3582 T12700 T12692 conj from,from
R3583 T12701 T12702 det the,effects
R3584 T12535 T12534 cc or,FIAU
R3585 T12702 T12700 pobj effects,from
R3586 T12703 T12702 prep of,effects
R3587 T12536 T12534 conj ganciclovir,FIAU
R3588 T12704 T12703 pobj p53GFP,of
R3589 T12705 T12673 punct .,done
R3590 T12537 T12508 punct .,attempted
R3591 T12707 T12708 nsubj We,observed
R3592 T12539 T12540 advmod Strikingly,identified
R3593 T12709 T12710 det a,cells
R3594 T12710 T12708 dobj cells,observed
R3595 T12541 T12540 punct ", ",identified
R3596 T12711 T12710 amod few,cells
R3597 T12542 T12543 det no,clones
R3598 T12712 T12710 amod fluorescent,cells
R3599 T12713 T12714 advmod only,when
R3600 T12714 T12715 advmod when,co-electroporated
R3601 T12715 T12710 advcl co-electroporated,cells
R3602 T12543 T12540 nsubjpass clones,identified
R3603 T12716 T12717 det the,plasmid
R3604 T12717 T12715 nsubjpass plasmid,co-electroporated
R3605 T12718 T12719 compound Cre,expression
R3606 T12544 T12543 prep with,clones
R3607 T12719 T12717 compound expression,plasmid
R3608 T12720 T12715 auxpass was,co-electroporated
R3609 T12721 T12715 punct ", ",co-electroporated
R3610 T12545 T12546 det an,allele
R3611 T12722 T12715 advcl suggesting,co-electroporated
R3612 T12723 T12724 mark that,resulted
R3613 T12724 T12722 ccomp resulted,suggesting
R3614 T12546 T12544 pobj allele,with
R3615 T12725 T12726 amod such,cells
R3616 T12726 T12724 nsubj cells,resulted
R3617 T12727 T12724 prep from,resulted
R3618 T12547 T12546 amod exchanged,allele
R3619 T12728 T12727 pobj RMCE,from
R3620 T12729 T12708 punct .,observed
R3621 T12548 T12540 auxpass were,identified
R3622 T12731 T12732 advmod Importantly,had
R3623 T12733 T12732 punct ", ",had
R3624 T12549 T12550 punct (,shown
R3625 T12734 T12735 det the,cells
R3626 T12735 T12732 nsubj cells,had
R3627 T12736 T12735 amod rare,cells
R3628 T12550 T12540 parataxis shown,identified
R3629 T12737 T12735 amod fluorescent,cells
R3630 T12738 T12739 det a,appearance
R3631 T12739 T12732 dobj appearance,had
R3632 T12551 T12550 nsubj data,shown
R3633 T12740 T12739 amod flat,appearance
R3634 T12741 T12739 punct ", ",appearance
R3635 T12742 T12739 punct ‘,appearance
R3636 T12552 T12550 neg not,shown
R3637 T12743 T12744 amod fried,egg
R3638 T12744 T12739 nmod egg,appearance
R3639 T12745 T12744 punct -,egg
R3640 T12553 T12550 punct ),shown
R3641 T12746 T12739 punct ’,appearance
R3642 T12747 T12739 amod typical,appearance
R3643 T12748 T12747 prep of,typical
R3644 T12554 T12540 punct .,identified
R3645 T12749 T12750 amod senescent,cells
R3646 T12750 T12748 pobj cells,of
R3647 T12751 T12752 punct (,5A
R3648 T12556 T12557 nsubj RMCE,showed
R3649 T12752 T12732 parataxis 5A,had
R3650 T12753 T12752 compound Figure,5A
R3651 T12754 T12752 punct ),5A
R3652 T12755 T12732 punct ", ",had
R3653 T12756 T12732 cc and,had
R3654 T12757 T12758 advmod when,observed
R3655 T12758 T12761 advcl observed,detached
R3656 T12558 T12556 prep with,RMCE
R3657 T12759 T12758 nsubjpass plates,observed
R3658 T12760 T12758 auxpass were,observed
R3659 T12761 T12732 conj detached,had
R3660 T12762 T12763 nummod 5,days
R3661 T12559 T12558 pobj p53GFP,with
R3662 T12763 T12764 npadvmod days,later
R3663 T12764 T12758 advmod later,observed
R3664 T12765 T12761 punct ", ",detached
R3665 T12560 T12556 prep in,RMCE
R3666 T12766 T12767 det the,cells
R3667 T12767 T12761 nsubj cells,detached
R3668 T12561 T12562 compound ES,cells
R3669 T12768 T12761 aux had,detached
R3670 T12769 T12761 punct .,detached
R3671 T12562 T12560 pobj cells,in
R3672 T12771 T12772 advmod Altogether,suggest
R3673 T12773 T12772 punct ", ",suggest
R3674 T12563 T12564 mark that,expressed
R3675 T12774 T12775 det the,results
R3676 T12775 T12772 nsubj results,suggest
R3677 T12776 T12777 mark that,give
R3678 T12564 T12557 ccomp expressed,showed
R3679 T12777 T12772 ccomp give,suggest
R3680 T12778 T12777 nsubj RMCE,give
R3681 T12779 T12777 aux can,give
R3682 T12565 T12564 nsubjpass p53GFP,expressed
R3683 T12780 T12777 dobj rise,give
R3684 T12781 T12777 prep to,give
R3685 T12782 T12783 npadvmod p53GFP,expressing
R3686 T12566 T12564 auxpass is,expressed
R3687 T12783 T12785 amod expressing,MEFs
R3688 T12784 T12783 punct -,expressing
R3689 T12567 T12564 prep at,expressed
R3690 T12785 T12781 pobj MEFs,to
R3691 T12786 T12777 punct ", ",give
R3692 T12787 T12777 cc but,give
R3693 T12788 T12789 nsubj they,die
R3694 T12568 T12569 amod high,levels
R3695 T12789 T12777 conj die,give
R3696 T12790 T12789 advmod rapidly,die
R3697 T12569 T12567 pobj levels,at
R3698 T12791 T12789 prep owing,die
R3699 T12792 T12791 prep to,owing
R3700 T12793 T12794 compound p53GFP,toxicity
R3701 T12570 T12571 punct (,3A
R3702 T12794 T12792 pobj toxicity,to
R3703 T12795 T12772 punct .,suggest
R3704 T12571 T12564 parataxis 3A,expressed
R3705 T12797 T12798 nsubj We,performed
R3706 T12799 T12798 advmod also,performed
R3707 T12572 T12571 compound Figure,3A
R3708 T12800 T12798 dobj RMCE,performed
R3709 T12573 T12571 punct ),3A
R3710 T12574 T12557 punct ", ",showed
R3711 T12801 T12798 prep in,performed
R3712 T12802 T12803 nmod p53RMCE,MEFs
R3713 T12575 T12557 cc and,showed
R3714 T12803 T12801 pobj MEFs,in
R3715 T12804 T12802 punct /,p53RMCE
R3716 T12576 T12577 mark as,mentioned
R3717 T12805 T12802 punct −,p53RMCE
R3718 T12806 T12798 prep with,performed
R3719 T12577 T12578 advcl mentioned,activated
R3720 T12807 T12808 det the,construct
R3721 T12808 T12806 pobj construct,with
R3722 T12578 T12557 conj activated,showed
R3723 T12809 T12808 compound p53ΔPGFP,construct
R3724 T12810 T12798 punct .,performed
R3725 T12579 T12577 advmod before,mentioned
R3726 T12812 T12813 npadvmod p53ΔPGFP,expressing
R3727 T12813 T12815 amod expressing,MEFs
R3728 T12814 T12813 punct -,expressing
R3729 T12580 T12578 punct ", ",activated
R3730 T12815 T12816 nsubj MEFs,were
R3731 T12581 T12582 det the,pathway
R3732 T12817 T12816 acomp viable,were
R3733 T12818 T12816 punct ", ",were
R3734 T12819 T12816 conj recovered,were
R3735 T12582 T12578 nsubjpass pathway,activated
R3736 T12820 T12819 prep with,recovered
R3737 T12821 T12822 det an,efficiency
R3738 T12583 T12582 compound p53,pathway
R3739 T12822 T12820 pobj efficiency,with
R3740 T12823 T12822 prep of,efficiency
R3741 T12824 T12825 punct ∼,40
R3742 T12584 T12585 dep that,activated
R3743 T12825 T12826 nummod 40,%
R3744 T12826 T12823 pobj %,of
R3745 T12827 T12819 punct ", ",recovered
R3746 T12585 T12582 relcl activated,pathway
R3747 T12828 T12819 cc and,recovered
R3748 T12829 T12819 punct ", ",recovered
R3749 T12830 T12831 mark as,expected
R3750 T12586 T12585 aux can,activated
R3751 T12831 T12832 advcl expected,expressed
R3752 T12832 T12819 conj expressed,recovered
R3753 T12833 T12831 prep from,expected
R3754 T12587 T12585 auxpass be,activated
R3755 T12834 T12835 compound ES,cell
R3756 T12835 T12836 compound cell,experiments
R3757 T12836 T12833 pobj experiments,from
R3758 T12837 T12832 punct ", ",expressed
R3759 T12838 T12839 det a,protein
R3760 T12588 T12585 prep in,activated
R3761 T12839 T12832 dobj protein,expressed
R3762 T12840 T12839 compound p53ΔPGFP,protein
R3763 T12841 T12832 prep at,expressed
R3764 T12842 T12843 advmod much,lower
R3765 T12906 T12878 aux did,arrest
R3766 T12843 T12844 amod lower,levels
R3767 T12844 T12841 pobj levels,at
R3768 T12845 T12844 prep than,levels
R3769 T12846 T12845 pobj p53WT,than
R3770 T12907 T12878 neg not,arrest
R3771 T12847 T12848 punct (,5B
R3772 T12848 T12832 parataxis 5B,expressed
R3773 T12849 T12848 compound Figure,5B
R3774 T12850 T12848 punct ),5B
R3775 T12908 T12909 nmod p53ΔPGFP,MEFs
R3776 T12851 T12816 punct .,were
R3777 T12909 T12878 dobj MEFs,arrest
R3778 T12853 T12854 prep Unlike,are
R3779 T12855 T12856 compound WT,MEFs
R3780 T12910 T12908 punct /,p53ΔPGFP
R3781 T12856 T12853 pobj MEFs,Unlike
R3782 T12857 T12854 punct ", ",are
R3783 T12858 T12859 compound p53ΔP,ΔP
R3784 T12911 T12908 punct −,p53ΔPGFP
R3785 T12859 T12861 compound ΔP,MEFs
R3786 T12860 T12859 punct /,ΔP
R3787 T12912 T12913 punct (,5C
R3788 T12861 T12854 nsubj MEFs,are
R3789 T12862 T12854 acomp unable,are
R3790 T12863 T12864 aux to,arrest
R3791 T12913 T12878 parataxis 5C,arrest
R3792 T12864 T12862 xcomp arrest,unable
R3793 T12865 T12864 dobj cycling,arrest
R3794 T12866 T12864 prep after,arrest
R3795 T12914 T12913 compound Figure,5C
R3796 T12867 T12866 pobj irradiation,after
R3797 T12868 T12869 punct (,5
R3798 T12869 T12854 parataxis 5,are
R3799 T12915 T12913 punct ),5C
R3800 T12870 T12869 punct ),5
R3801 T12871 T12854 punct .,are
R3802 T12916 T12874 punct .,found
R3803 T12873 T12874 advmod Likewise,found
R3804 T12918 T12919 det These,data
R3805 T12875 T12874 punct ", ",found
R3806 T12876 T12874 nsubj we,found
R3807 T12877 T12878 mark that,arrest
R3808 T12919 T12920 nsubj data,show
R3809 T12878 T12874 ccomp arrest,found
R3810 T12921 T12922 mark that,enables
R3811 T12879 T12880 compound irradiation,doses
R3812 T12880 T12878 nsubj doses,arrest
R3813 T12881 T12882 dep that,arrested
R3814 T12882 T12880 relcl arrested,doses
R3815 T12922 T12920 ccomp enables,show
R3816 T12883 T12884 nmod p53RMCE,MEFs
R3817 T12884 T12882 dobj MEFs,arrested
R3818 T12923 T12924 det a,reaction
R3819 T12885 T12883 punct /,p53RMCE
R3820 T12886 T12883 punct −,p53RMCE
R3821 T12887 T12880 punct (,doses
R3822 T12924 T12922 nsubj reaction,enables
R3823 T12888 T12889 dep which,express
R3824 T12889 T12880 relcl express,doses
R3825 T12890 T12891 det a,p53
R3826 T12925 T12924 amod single,reaction
R3827 T12891 T12889 dobj p53,express
R3828 T12892 T12893 amod wild,type
R3829 T12926 T12927 compound RMCE,ASAP
R3830 T12893 T12891 compound type,p53
R3831 T12894 T12893 punct -,type
R3832 T12895 T12891 prep from,p53
R3833 T12927 T12924 compound ASAP,reaction
R3834 T12896 T12897 det the,locus
R3835 T12897 T12895 pobj locus,from
R3836 T12898 T12899 npadvmod RMCE,ready
R3837 T12928 T12927 punct -,ASAP
R3838 T12899 T12897 amod ready,locus
R3839 T12900 T12899 punct -,ready
R3840 T12901 T12878 punct ", ",arrest
R3841 T12929 T12924 prep in,reaction
R3842 T12902 T12878 parataxis see,arrest
R3843 T12903 T12902 dobj Figure,see
R3844 T12930 T12931 amod heterozygous,MEFs
R3845 T12904 T12903 nummod 2,Figure
R3846 T12905 T12878 punct ),arrest
R3847 T12931 T12929 pobj MEFs,in
R3848 T12932 T12922 dobj detection,enables
R3849 T12933 T12932 prep of,detection
R3850 T13012 T13013 prep in,accelerate
R3851 T12934 T12935 det a,phenotype
R3852 T13013 T13001 conj accelerate,enables
R3853 T13014 T13015 amod heterozygous,MEFs
R3854 T12935 T12933 pobj phenotype,of
R3855 T13015 T13012 pobj MEFs,in
R3856 T13016 T13013 aux to,accelerate
R3857 T13017 T13018 amod phenotypic,analyses
R3858 T12936 T12935 amod recessive,phenotype
R3859 T13018 T13013 dobj analyses,accelerate
R3860 T13019 T12990 punct .,report
R3861 T13021 T13022 det The,success
R3862 T13022 T13023 nsubj success,relied
R3863 T12937 T12920 punct .,show
R3864 T13024 T13022 prep of,success
R3865 T13025 T13026 compound RMCE,ASAP
R3866 T12939 T12940 det The,results
R3867 T13026 T13024 pobj ASAP,of
R3868 T13027 T13026 punct -,ASAP
R3869 T13028 T13023 prep on,relied
R3870 T13029 T13030 det the,use
R3871 T13030 T13028 pobj use,on
R3872 T12940 T12941 nsubj results,confirm
R3873 T13031 T13030 amod integrated,use
R3874 T13032 T13030 prep of,use
R3875 T13033 T13034 amod inverted,sites
R3876 T12942 T12943 mark that,leads
R3877 T13034 T13032 pobj sites,of
R3878 T13035 T13036 amod heterologous,loxP
R3879 T13036 T13034 compound loxP,sites
R3880 T13037 T13034 punct ", ",sites
R3881 T12943 T12941 ccomp leads,confirm
R3882 T13038 T13039 det a,marker
R3883 T13039 T13034 conj marker,sites
R3884 T12944 T12943 csubj deleting,leads
R3885 T13040 T13041 amod positive,negative
R3886 T13041 T13043 nmod negative,selection
R3887 T13042 T13041 punct /,negative
R3888 T12945 T12946 det the,domain
R3889 T13043 T13039 compound selection,marker
R3890 T13044 T13045 dep that,preserves
R3891 T13045 T13039 relcl preserves,marker
R3892 T13046 T13047 det the,capacity
R3893 T12946 T12944 dobj domain,deleting
R3894 T13047 T13045 dobj capacity,preserves
R3895 T13048 T13047 compound germline,capacity
R3896 T13049 T13047 prep of,capacity
R3897 T12947 T12948 npadvmod proline,rich
R3898 T13050 T13051 compound ES,cells
R3899 T13051 T13049 pobj cells,of
R3900 T13052 T13039 punct ", ",marker
R3901 T12948 T12946 amod rich,domain
R3902 T13053 T13039 cc and,marker
R3903 T13054 T13039 punct ", ",marker
R3904 T12949 T12943 prep to,leads
R3905 T12950 T12951 advmod less,active
R3906 T13055 T13056 prep for,existence
R3907 T13056 T13039 conj existence,marker
R3908 T13057 T13058 amod somatic,cells
R3909 T12951 T12952 amod active,p53
R3910 T13058 T13055 pobj cells,for
R3911 T13059 T13056 punct ", ",existence
R3912 T13060 T13056 det the,existence
R3913 T13061 T13056 prep of,existence
R3914 T13062 T13063 det a,allele
R3915 T13063 T13061 pobj allele,of
R3916 T12952 T12949 pobj p53,to
R3917 T13064 T13063 amod knock,allele
R3918 T13065 T13064 punct -,knock
R3919 T13066 T13064 prt out,knock
R3920 T12953 T12952 prep with,p53
R3921 T13067 T13063 prep of,allele
R3922 T13068 T13069 det the,gene
R3923 T13069 T13067 pobj gene,of
R3924 T12954 T12955 amod impaired,control
R3925 T13070 T13069 prep of,gene
R3926 T13071 T13070 pobj interest,of
R3927 T12955 T12953 pobj control,with
R3928 T13072 T13023 punct .,relied
R3929 T13074 T13075 det These,characteristics
R3930 T12956 T12957 compound cell,cycle
R3931 T13075 T13076 nsubj characteristics,make
R3932 T13077 T13076 aux should,make
R3933 T12957 T12955 compound cycle,control
R3934 T13078 T13079 compound RMCE,ASAP
R3935 T13079 T13081 nsubj ASAP,technology
R3936 T13080 T13079 punct -,ASAP
R3937 T12958 T12941 punct ", ",confirm
R3938 T13081 T13076 ccomp technology,make
R3939 T13082 T13081 det a,technology
R3940 T13083 T13081 amod robust,technology
R3941 T12959 T12941 cc and,confirm
R3942 T13084 T13083 cc and,robust
R3943 T13085 T13083 conj general,robust
R3944 T13086 T13081 prep for,technology
R3945 T12960 T12961 advmod also,indicate
R3946 T13087 T13086 pobj analysis,for
R3947 T13088 T13087 prep of,analysis
R3948 T13089 T13090 amod mammalian,genes
R3949 T12961 T12941 conj indicate,confirm
R3950 T13090 T13088 pobj genes,of
R3951 T13091 T13087 prep under,analysis
R3952 T12962 T12963 mark that,alter
R3953 T13092 T13091 pobj conditions,under
R3954 T13093 T13094 dep that,preserve
R3955 T13094 T13092 relcl preserve,conditions
R3956 T12963 T12961 ccomp alter,indicate
R3957 T13095 T13096 amod normal,mechanisms
R3958 T13096 T13094 dobj mechanisms,preserve
R3959 T13097 T13096 compound control,mechanisms
R3960 T13098 T13094 prep in,preserve
R3961 T12964 T12965 det a,fusion
R3962 T13099 T13100 amod different,tissues
R3963 T13100 T13098 pobj tissues,in
R3964 T12965 T12963 nsubj fusion,alter
R3965 T13101 T13076 punct .,make
R3966 T12966 T12965 nmod GFP,fusion
R3967 T13103 T13104 prep In,used
R3968 T12967 T12965 nmod C,fusion
R3969 T13105 T13103 pobj addition,In
R3970 T13106 T13104 punct ", ",used
R3971 T13107 T13108 compound RMCE,ASAP
R3972 T12968 T12967 punct -,C
R3973 T13108 T13104 nsubjpass ASAP,used
R3974 T13109 T13108 punct -,ASAP
R3975 T12969 T12967 amod terminal,C
R3976 T13110 T13104 aux could,used
R3977 T13111 T13104 auxpass be,used
R3978 T13112 T13113 aux to,generate
R3979 T12970 T12963 aux can,alter
R3980 T13113 T13104 advcl generate,used
R3981 T13114 T13115 amod fibroblastic,lines
R3982 T13115 T13113 dobj lines,generate
R3983 T12971 T12963 advmod dramatically,alter
R3984 T13116 T13115 compound cell,lines
R3985 T13117 T13115 acl tailored,lines
R3986 T12972 T12973 compound p53,regulation
R3987 T12973 T12963 dobj regulation,alter
R3988 T13118 T13117 prep for,tailored
R3989 T12974 T12941 punct .,confirm
R3990 T13119 T13120 det the,targeting
R3991 T13120 T13118 pobj targeting,for
R3992 T13121 T13120 amod repeated,targeting
R3993 T12976 T12977 det A,summary
R3994 T13122 T13120 prep of,targeting
R3995 T13123 T13124 advmod widely,studied
R3996 T13124 T13125 amod studied,genes
R3997 T12977 T12978 nsubjpass summary,presented
R3998 T13125 T13122 pobj genes,of
R3999 T13126 T13125 punct (,genes
R4000 T13127 T13125 appos p53,genes
R4001 T12979 T12977 prep of,summary
R4002 T13128 T13127 punct ", ",p53
R4003 T13129 T13130 compound c,myc
R4004 T13130 T13127 conj myc,p53
R4005 T12980 T12981 poss our,results
R4006 T13131 T13130 punct -,myc
R4007 T13132 T13130 punct ", ",myc
R4008 T13133 T13134 compound NF,KB
R4009 T12981 T12979 pobj results,of
R4010 T13134 T13130 conj KB,myc
R4011 T13135 T13134 punct -,KB
R4012 T13136 T13134 punct ", ",KB
R4013 T12982 T12978 auxpass is,presented
R4014 T13137 T13134 conj etc.,KB
R4015 T13138 T13104 punct ),used
R4016 T13139 T13104 punct .,used
R4017 T12983 T12978 prep in,presented
R4018 T12984 T12983 pobj Table,in
R4019 T12985 T12984 nummod 1,Table
R4020 T12986 T12978 punct .,presented
R4021 T12988 T12989 det These,data
R4022 T12989 T12990 nsubj data,report
R4023 T12991 T12992 det the,development
R4024 T12992 T12990 dobj development,report
R4025 T12993 T12992 cc and,development
R4026 T12994 T12992 conj implementation,development
R4027 T12995 T12992 prep of,development
R4028 T12996 T12997 det an,approach
R4029 T12997 T12995 pobj approach,of
R4030 T12998 T12999 amod improved,RMCE
R4031 T12999 T12997 compound RMCE,approach
R4032 T13000 T13001 dep that,enables
R4033 T13001 T12997 relcl enables,approach
R4034 T13002 T13003 amod efficient,modification
R4035 T13003 T13001 dobj modification,enables
R4036 T13004 T13003 compound allele,modification
R4037 T13005 T13001 prep in,enables
R4038 T13006 T13007 compound ES,cells
R4039 T13007 T13005 pobj cells,in
R4040 T13008 T13009 aux to,generate
R4041 T13009 T13001 advcl generate,enables
R4042 T13010 T13009 dobj mice,generate
R4043 T13011 T13001 cc and,enables