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the human gene, the mouse Atrx gene is also X-linked, such that a direct disruption of the single Atrx allele in male ES cells would immediately give rise to the null state. No targeted clones were recovered after attempted homologous recombination in male E14TG2a ES cells using two different vectors that removed exon 18 of the Atrx gene. Exon 18 encodes the first of the seven motifs composing the conserved SNF2-like domain of Atrx (Figure 1); mutation of the corresponding motif of the yeast SNF2 protein has been shown to severely impair SWI/SNF-dependent gene expression [12]. The failure to recover targeted clones with these vectors suggested that Atrx may be important for normal ES cell growth and expansion and that direct targeting of the single locus may not be possible. We therefore adopted a conditional strategy for targeting exon 18 (Figure 2) and recovered two clones in which exon 18 has been flanked by loxP recognition sites for the Cre recombinase (Atrx flox allele in Figure 2A) (Figure 2B). This allele also contains a loxP-flanked MC1-neor cassette in intron 17 (Figure 2A). Northern and Western blot analyses (Figure 2D and 2E) confirmed that the Atrx flox clones continued to express both full-length Atrx protein and the truncated Atrxt isoform."}

{"project":"craft-ca-core-ex-dev","denotations":[{"id":"T1465","span":{"begin":9,"end":14},"obj":"NCBITaxon:9606"},{"id":"T1466","span":{"begin":15,"end":19},"obj":"SO_EXT:0000704"},{"id":"T1467","span":{"begin":25,"end":30},"obj":"NCBITaxon:10088"},{"id":"T1468","span":{"begin":31,"end":35},"obj":"PR_EXT:000004503"},{"id":"T1469","span":{"begin":36,"end":40},"obj":"SO_EXT:0000704"},{"id":"T1470","span":{"begin":49,"end":50},"obj":"GO:0000805"},{"id":"T1471","span":{"begin":103,"end":107},"obj":"PR_EXT:000004503"},{"id":"T1472","span":{"begin":108,"end":114},"obj":"SO_EXT:0001023"},{"id":"T1473","span":{"begin":118,"end":122},"obj":"PATO_UBERON_EXT:male_or_bearer_of_maleness"},{"id":"T1474","span":{"begin":123,"end":131},"obj":"CL:0002322"},{"id":"T1475","span":{"begin":126,"end":131},"obj":"CL_GO_EXT:cell"},{"id":"T1476","span":{"begin":167,"end":171},"obj":"SO_EXT:sequence_nullness"},{"id":"T1477","span":{"begin":191,"end":197},"obj":"SO_EXT:sequence_cloned_entity"},{"id":"T1478","span":{"begin":240,"end":253},"obj":"GO_SO_EXT:sequence_rearrangement_process"},{"id":"T1479","span":{"begin":257,"end":261},"obj":"PATO_UBERON_EXT:male_or_bearer_of_maleness"},{"id":"T1480","span":{"begin":270,"end":278},"obj":"CL:0002322"},{"id":"T1481","span":{"begin":273,"end":278},"obj":"CL_GO_EXT:cell"},{"id":"T1482","span":{"begin":299,"end":306},"obj":"SO_EXT:0000440"},{"id":"T1483","span":{"begin":320,"end":324},"obj":"SO_EXT:0000147"},{"id":"T1484","span":{"begin":335,"end":339},"obj":"PR_EXT:000004503"},{"id":"T1485","span":{"begin":340,"end":344},"obj":"SO_EXT:0000704"},{"id":"T1486","span":{"begin":346,"end":350},"obj":"SO_EXT:0000147"},{"id":"T1487","span":{"begin":354,"end":361},"obj":"SO_EXT:sequence_coding_function"},{"id":"T1488","span":{"begin":385,"end":391},"obj":"SO_EXT:sequence_or_structure_motif"},{"id":"T1489","span":{"begin":406,"end":415},"obj":"SO_EXT:biological_conservation_process_or_quality"},{"id":"T1490","span":{"begin":416,"end":420},"obj":"PR_EXT:P22082"},{"id":"T1491","span":{"begin":426,"end":432},"obj":"SO_EXT:0000417"},{"id":"T1492","span":{"begin":436,"end":440},"obj":"PR_EXT:000004503"},{"id":"T1493","span":{"begin":453,"end":461},"obj":"SO_EXT:sequence_alteration_entity_or_process"},{"id":"T1494","span":{"begin":483,"end":488},"obj":"SO_EXT:sequence_or_structure_motif"},{"id":"T1495","span":{"begin":496,"end":501},"obj":"NCBITaxon_EXT:yeast"},{"id":"T1496","span":{"begin":502,"end":506},"obj":"PR_EXT:P22082"},{"id":"T1497","span":{"begin":507,"end":514},"obj":"CHEBI_PR_EXT:protein"},{"id":"T1498","span":{"begin":549,"end":556},"obj":"GO:0016514"},{"id":"T1499","span":{"begin":567,"end":571},"obj":"SO_EXT:0000704"},{"id":"T1500","span":{"begin":567,"end":582},"obj":"GO:0010467"},{"id":"T1501","span":{"begin":621,"end":627},"obj":"SO_EXT:sequence_cloned_entity"},{"id":"T1502","span":{"begin":639,"end":646},"obj":"SO_EXT:0000440"},{"id":"T1503","span":{"begin":662,"end":666},"obj":"PR_EXT:000004503"},{"id":"T1504","span":{"begin":695,"end":702},"obj":"CL:0002322"},{"id":"T1505","span":{"begin":698,"end":702},"obj":"CL_GO_EXT:cell"},{"id":"T1506","span":{"begin":703,"end":709},"obj":"GO_EXT:biological_growth_entity_or_process"},{"id":"T1507","span":{"begin":849,"end":853},"obj":"SO_EXT:0000147"},{"id":"T1508","span":{"begin":886,"end":892},"obj":"SO_EXT:sequence_cloned_entity"},{"id":"T1509","span":{"begin":902,"end":906},"obj":"SO_EXT:0000147"},{"id":"T1510","span":{"begin":919,"end":926},"obj":"SO:0000357"},{"id":"T1511","span":{"begin":930,"end":952},"obj":"SO_EXT:0000346"},{"id":"T1512","span":{"begin":965,"end":976},"obj":"GO_EXT:recombinase"},{"id":"T1513","span":{"begin":978,"end":982},"obj":"PR_EXT:000004503"},{"id":"T1514","span":{"begin":983,"end":987},"obj":"SO:0000359"},{"id":"T1515","span":{"begin":988,"end":994},"obj":"SO_EXT:0001023"},{"id":"T1516","span":{"begin":1027,"end":1033},"obj":"SO_EXT:0001023"},{"id":"T1517","span":{"begin":1050,"end":1062},"obj":"SO:0000359"},{"id":"T1518","span":{"begin":1067,"end":1071},"obj":"GO_EXT:0008910"},{"id":"T1519","span":{"begin":1072,"end":1080},"obj":"SO_EXT:0005853"},{"id":"T1520","span":{"begin":1084,"end":1090},"obj":"SO_EXT:0000188"},{"id":"T1521","span":{"begin":1180,"end":1184},"obj":"PR_EXT:000004503"},{"id":"T1522","span":{"begin":1185,"end":1189},"obj":"SO:0000359"},{"id":"T1523","span":{"begin":1190,"end":1196},"obj":"SO_EXT:sequence_cloned_entity"},{"id":"T1524","span":{"begin":1210,"end":1217},"obj":"GO:0010467"},{"id":"T1525","span":{"begin":1235,"end":1239},"obj":"PR_EXT:000004503"},{"id":"T1526","span":{"begin":1240,"end":1247},"obj":"CHEBI_PR_EXT:protein"},{"id":"T1527","span":{"begin":1256,"end":1265},"obj":"SO_EXT:sequence_truncation_process"},{"id":"T1528","span":{"begin":1272,"end":1279},"obj":"SO_EXT:0001060"}],"text":"Like the human gene, the mouse Atrx gene is also X-linked, such that a direct disruption of the single Atrx allele in male ES cells would immediately give rise to the null state. No targeted clones were recovered after attempted homologous recombination in male E14TG2a ES cells using two different vectors that removed exon 18 of the Atrx gene. Exon 18 encodes the first of the seven motifs composing the conserved SNF2-like domain of Atrx (Figure 1); mutation of the corresponding motif of the yeast SNF2 protein has been shown to severely impair SWI/SNF-dependent gene expression [12]. The failure to recover targeted clones with these vectors suggested that Atrx may be important for normal ES cell growth and expansion and that direct targeting of the single locus may not be possible. We therefore adopted a conditional strategy for targeting exon 18 (Figure 2) and recovered two clones in which exon 18 has been flanked by loxP recognition sites for the Cre recombinase (Atrx flox allele in Figure 2A) (Figure 2B). This allele also contains a loxP-flanked MC1-neor cassette in intron 17 (Figure 2A). Northern and Western blot analyses (Figure 2D and 2E) confirmed that the Atrx flox clones continued to express both full-length Atrx protein and the truncated Atrxt isoform."}

{"project":"2_test","denotations":[{"id":"16628246-8871545-85799746","span":{"begin":584,"end":586},"obj":"8871545"},{"id":"T3548","span":{"begin":584,"end":586},"obj":"8871545"}],"text":"Like the human gene, the mouse Atrx gene is also X-linked, such that a direct disruption of the single Atrx allele in male ES cells would immediately give rise to the null state. No targeted clones were recovered after attempted homologous recombination in male E14TG2a ES cells using two different vectors that removed exon 18 of the Atrx gene. Exon 18 encodes the first of the seven motifs composing the conserved SNF2-like domain of Atrx (Figure 1); mutation of the corresponding motif of the yeast SNF2 protein has been shown to severely impair SWI/SNF-dependent gene expression [12]. The failure to recover targeted clones with these vectors suggested that Atrx may be important for normal ES cell growth and expansion and that direct targeting of the single locus may not be possible. We therefore adopted a conditional strategy for targeting exon 18 (Figure 2) and recovered two clones in which exon 18 has been flanked by loxP recognition sites for the Cre recombinase (Atrx flox allele in Figure 2A) (Figure 2B). This allele also contains a loxP-flanked MC1-neor cassette in intron 17 (Figure 2A). Northern and Western blot analyses (Figure 2D and 2E) confirmed that the Atrx flox clones continued to express both full-length Atrx protein and the truncated Atrxt isoform."}

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