Utility of C57BL/6J x 129/SvJae embryonic stem cells for generating chromosomal deletions:: tolerance to γ radiation and microsatellite polymorphism

被引:20
作者
You, Y
Bersgtram, R
Klemm, M
Nelson, H
Jaenisch, R
Schimenti, J
机构
[1] Jackson Lab, Bar Harbor, ME 04609 USA
[2] Whitehead Inst Biomed Res, Cambridge Ctr 9, Cambridge, MA 02142 USA
关键词
D O I
10.1007/s003359900731
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
We have previously reported a method for making nested deletion complexes in mice by irradiation of ES cells. The kev to this technology is that F-1 hybrid ES cells (called v17.2) of the genotype (BALB/cTa x 129/Sv(Jae)) retain germline colonizing ability after exposure to levels of ionizing radiation that induce chromosomal deletions. In an effort to identify other genotypes of ES cells that are suitable for this technology, the radiation sensitivity of the cell line v6.4, which is of the genotype (C57BL/6J x radiation exposures, the developmental potential of these cells was assayed by injecting them into blastocysts to generate chimeric mice. These experiments showed that while cell lethality increased as the level of radiation increased, the surviving ES cells retained full totipotency at all exposure levels, up to 400 Rads. Because polymorphism between parental microsatellite alleles in the F-1 hyrbrid ES cells is important for ascertaining the sizes of induced deletions, the 129/SvJ and 129/Sv(Jae) allele sizes of 38 microsatellite loci on chromosome (Chr) 17 were determined. This revealed a higher level of polymorphism between 129 and C57BL/6J on Chr 17. The radiation tolerance, high polymorphism between parental strains, and presence of the widely used C57BL/6J strain component make v6.3 ES cells an attractive cell line for generating radiation-induced chromosomal deletions.
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页码:232 / 234
页数:3
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