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generate the full deletion in ES cells, the Atrx flox clones (1/F12 and 1/G11) were transiently transfected with a Cre-recombinase expression plasmid (pCAGGS-Cre-IRESpuro), and subclones were recovered bearing an allele (Atrx Δ18Δneo in Figure 2A) in which both exon 18 and the neor cassette had been deleted by the Cre recombinase (resulting from the recombination event labelled “C” in the Atrx flox allele shown in Figure 2A) (Figure 2C). Northern and Western blot analyses (Figure 2D and 2E) revealed that the full-length Atrx transcript and protein is completely abolished in the Atrx Δ18Δneo recombinant clones, suggesting that deletion of this region has a highly destabilising effect on the full-length transcript. As expected, the truncated Atrxt isoform, the transcript of which is terminated within intron 11 [4], is unaffected by the deletion of exon 18 (Figure 2E). While the function of Atrxt is not yet clear, this isoform, which contains the PHD-like domain but not the SWI/SNF motifs (Figure 1), is unlikely to be functionally equivalent to the full-length protein. Thus, a conditional knockout strategy allowed the isolation of ES cells that are null for full-length Atrx."}

{"project":"craft-ca-core-ex-dev","denotations":[{"id":"T1529","span":{"begin":21,"end":29},"obj":"SO_EXT:sequence_deletion_entity_or_process"},{"id":"T1530","span":{"begin":33,"end":41},"obj":"CL:0002322"},{"id":"T1531","span":{"begin":36,"end":41},"obj":"CL_GO_EXT:cell"},{"id":"T1532","span":{"begin":47,"end":51},"obj":"PR_EXT:000004503"},{"id":"T1533","span":{"begin":52,"end":56},"obj":"SO:0000359"},{"id":"T1534","span":{"begin":57,"end":63},"obj":"SO_EXT:sequence_cloned_entity"},{"id":"T1535","span":{"begin":99,"end":110},"obj":"GO:0009294"},{"id":"T1536","span":{"begin":122,"end":133},"obj":"GO_EXT:recombinase"},{"id":"T1537","span":{"begin":134,"end":144},"obj":"GO:0010467"},{"id":"T1538","span":{"begin":145,"end":152},"obj":"SO_EXT:0000155"},{"id":"T1539","span":{"begin":165,"end":169},"obj":"SO_EXT:0000243"},{"id":"T1540","span":{"begin":216,"end":222},"obj":"SO_EXT:0001023"},{"id":"T1541","span":{"begin":224,"end":228},"obj":"PR_EXT:000004503"},{"id":"T1542","span":{"begin":265,"end":269},"obj":"SO_EXT:0000147"},{"id":"T1543","span":{"begin":281,"end":285},"obj":"GO_EXT:0008910"},{"id":"T1544","span":{"begin":286,"end":294},"obj":"SO_EXT:0005853"},{"id":"T1545","span":{"begin":304,"end":311},"obj":"SO_EXT:sequence_deletion_process"},{"id":"T1546","span":{"begin":323,"end":334},"obj":"GO_EXT:recombinase"},{"id":"T1547","span":{"begin":355,"end":368},"obj":"GO_SO_EXT:sequence_rearrangement_process"},{"id":"T1548","span":{"begin":395,"end":399},"obj":"PR_EXT:000004503"},{"id":"T1549","span":{"begin":400,"end":404},"obj":"SO:0000359"},{"id":"T1550","span":{"begin":405,"end":411},"obj":"SO_EXT:0001023"},{"id":"T1551","span":{"begin":529,"end":533},"obj":"PR_EXT:000004503"},{"id":"T1552","span":{"begin":534,"end":544},"obj":"SO_EXT:0000673"},{"id":"T1553","span":{"begin":549,"end":556},"obj":"CHEBI_PR_EXT:protein"},{"id":"T1554","span":{"begin":588,"end":592},"obj":"PR_EXT:000004503"},{"id":"T1555","span":{"begin":601,"end":612},"obj":"GO_SO_EXT:sequence_rearrangement_process"},{"id":"T1556","span":{"begin":613,"end":619},"obj":"SO_EXT:sequence_cloned_entity"},{"id":"T1557","span":{"begin":637,"end":645},"obj":"SO_EXT:sequence_deletion_entity_or_process"},{"id":"T1558","span":{"begin":714,"end":724},"obj":"SO_EXT:0000673"},{"id":"T1559","span":{"begin":743,"end":752},"obj":"SO_EXT:sequence_truncation_process"},{"id":"T1560","span":{"begin":759,"end":766},"obj":"SO_EXT:0001060"},{"id":"T1561","span":{"begin":772,"end":782},"obj":"SO_EXT:0000673"},{"id":"T1562","span":{"begin":813,"end":819},"obj":"SO_EXT:0000188"},{"id":"T1563","span":{"begin":849,"end":857},"obj":"SO_EXT:sequence_deletion_entity_or_process"},{"id":"T1564","span":{"begin":861,"end":865},"obj":"SO_EXT:0000147"},{"id":"T1565","span":{"begin":933,"end":940},"obj":"SO_EXT:0001060"},{"id":"T1566","span":{"begin":970,"end":976},"obj":"SO_EXT:0000417"},{"id":"T1567","span":{"begin":989,"end":996},"obj":"GO:0016514"},{"id":"T1568","span":{"begin":997,"end":1003},"obj":"SO_EXT:sequence_or_structure_motif"},{"id":"T1569","span":{"begin":1077,"end":1084},"obj":"CHEBI_PR_EXT:protein"},{"id":"T1570","span":{"begin":1149,"end":1157},"obj":"CL:0002322"},{"id":"T1571","span":{"begin":1152,"end":1157},"obj":"CL_GO_EXT:cell"},{"id":"T1572","span":{"begin":1167,"end":1171},"obj":"SO_EXT:sequence_nullness"},{"id":"T1573","span":{"begin":1188,"end":1192},"obj":"PR_EXT:000004503"}],"text":"To generate the full deletion in ES cells, the Atrx flox clones (1/F12 and 1/G11) were transiently transfected with a Cre-recombinase expression plasmid (pCAGGS-Cre-IRESpuro), and subclones were recovered bearing an allele (Atrx Δ18Δneo in Figure 2A) in which both exon 18 and the neor cassette had been deleted by the Cre recombinase (resulting from the recombination event labelled “C” in the Atrx flox allele shown in Figure 2A) (Figure 2C). Northern and Western blot analyses (Figure 2D and 2E) revealed that the full-length Atrx transcript and protein is completely abolished in the Atrx Δ18Δneo recombinant clones, suggesting that deletion of this region has a highly destabilising effect on the full-length transcript. As expected, the truncated Atrxt isoform, the transcript of which is terminated within intron 11 [4], is unaffected by the deletion of exon 18 (Figure 2E). While the function of Atrxt is not yet clear, this isoform, which contains the PHD-like domain but not the SWI/SNF motifs (Figure 1), is unlikely to be functionally equivalent to the full-length protein. Thus, a conditional knockout strategy allowed the isolation of ES cells that are null for full-length Atrx."}

{"project":"2_test","denotations":[{"id":"16628246-14729260-85799747","span":{"begin":824,"end":825},"obj":"14729260"},{"id":"T20654","span":{"begin":824,"end":825},"obj":"14729260"}],"text":"To generate the full deletion in ES cells, the Atrx flox clones (1/F12 and 1/G11) were transiently transfected with a Cre-recombinase expression plasmid (pCAGGS-Cre-IRESpuro), and subclones were recovered bearing an allele (Atrx Δ18Δneo in Figure 2A) in which both exon 18 and the neor cassette had been deleted by the Cre recombinase (resulting from the recombination event labelled “C” in the Atrx flox allele shown in Figure 2A) (Figure 2C). Northern and Western blot analyses (Figure 2D and 2E) revealed that the full-length Atrx transcript and protein is completely abolished in the Atrx Δ18Δneo recombinant clones, suggesting that deletion of this region has a highly destabilising effect on the full-length transcript. As expected, the truncated Atrxt isoform, the transcript of which is terminated within intron 11 [4], is unaffected by the deletion of exon 18 (Figure 2E). While the function of Atrxt is not yet clear, this isoform, which contains the PHD-like domain but not the SWI/SNF motifs (Figure 1), is unlikely to be functionally equivalent to the full-length protein. Thus, a conditional knockout strategy allowed the isolation of ES cells that are null for full-length Atrx."}

{"project":"craft-ca-core-dev","denotations":[{"id":"T1433","span":{"begin":33,"end":41},"obj":"CL:0002322"},{"id":"T1434","span":{"begin":47,"end":51},"obj":"PR:000004503"},{"id":"T1435","span":{"begin":52,"end":56},"obj":"SO:0000359"},{"id":"T1436","span":{"begin":99,"end":110},"obj":"GO:0009294"},{"id":"T1437","span":{"begin":134,"end":144},"obj":"GO:0010467"},{"id":"T1438","span":{"begin":145,"end":152},"obj":"SO:0000155"},{"id":"T1439","span":{"begin":165,"end":169},"obj":"SO:0000243"},{"id":"T1440","span":{"begin":216,"end":222},"obj":"SO:0001023"},{"id":"T1441","span":{"begin":224,"end":228},"obj":"PR:000004503"},{"id":"T1442","span":{"begin":265,"end":269},"obj":"SO:0000147"},{"id":"T1443","span":{"begin":286,"end":294},"obj":"SO:0005853"},{"id":"T1444","span":{"begin":395,"end":399},"obj":"PR:000004503"},{"id":"T1445","span":{"begin":400,"end":404},"obj":"SO:0000359"},{"id":"T1446","span":{"begin":405,"end":411},"obj":"SO:0001023"},{"id":"T1447","span":{"begin":529,"end":533},"obj":"PR:000004503"},{"id":"T1448","span":{"begin":534,"end":544},"obj":"SO:0000673"},{"id":"T1449","span":{"begin":588,"end":592},"obj":"PR:000004503"},{"id":"T1450","span":{"begin":714,"end":724},"obj":"SO:0000673"},{"id":"T1451","span":{"begin":759,"end":766},"obj":"SO:0001060"},{"id":"T1452","span":{"begin":772,"end":782},"obj":"SO:0000673"},{"id":"T1453","span":{"begin":813,"end":819},"obj":"SO:0000188"},{"id":"T1454","span":{"begin":861,"end":865},"obj":"SO:0000147"},{"id":"T1455","span":{"begin":933,"end":940},"obj":"SO:0001060"},{"id":"T1456","span":{"begin":970,"end":976},"obj":"SO:0000417"},{"id":"T1457","span":{"begin":989,"end":996},"obj":"GO:0016514"},{"id":"T1458","span":{"begin":1149,"end":1157},"obj":"CL:0002322"},{"id":"T1459","span":{"begin":1188,"end":1192},"obj":"PR:000004503"}],"text":"To generate the full deletion in ES cells, the Atrx flox clones (1/F12 and 1/G11) were transiently transfected with a Cre-recombinase expression plasmid (pCAGGS-Cre-IRESpuro), and subclones were recovered bearing an allele (Atrx Δ18Δneo in Figure 2A) in which both exon 18 and the neor cassette had been deleted by the Cre recombinase (resulting from the recombination event labelled “C” in the Atrx flox allele shown in Figure 2A) (Figure 2C). Northern and Western blot analyses (Figure 2D and 2E) revealed that the full-length Atrx transcript and protein is completely abolished in the Atrx Δ18Δneo recombinant clones, suggesting that deletion of this region has a highly destabilising effect on the full-length transcript. As expected, the truncated Atrxt isoform, the transcript of which is terminated within intron 11 [4], is unaffected by the deletion of exon 18 (Figure 2E). While the function of Atrxt is not yet clear, this isoform, which contains the PHD-like domain but not the SWI/SNF motifs (Figure 1), is unlikely to be functionally equivalent to the full-length protein. Thus, a conditional knockout strategy allowed the isolation of ES cells that are null for full-length Atrx."}