Utility of C57BL/6J x 129/SvJae embryonic stem cells for generating chromosomal deletions:: tolerance to γ radiation and microsatellite polymorphism
Utility of C57BL/6J x 129/SvJae embryonic stem cells for generating chromosomal deletions:: tolerance to γ radiation and microsatellite polymorphism
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DOI：
10.1007/s003359900731
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发表时间：
1998-03-01
期刊：
影响因子：
2.5
通讯作者：
Schimenti, J
中科院分区：
文献类型：
--
作者：
You, Y;Bersgtram, R;Schimenti, J
我们以前曾报道过一种通过照射ES细胞在小鼠体内制造嵌套缺失复合体的方法。这项技术的关键是，BALB/CTA x 129/Sv(JAE)基因的F-1杂交ES细胞(称为v17.2)在暴露于导致染色体缺失的电离辐射水平后，仍保持着生殖系定植能力。为了鉴定适合这项技术的ES细胞的其他基因类型，通过将这些细胞注射到囊胚中产生嵌合体小鼠来检测这些细胞的辐射敏感性，该细胞系属于(C57BL/6Jx)辐射暴露的基因。这些实验表明，虽然细胞致死率随着辐射水平的增加而增加，但存活的ES细胞在所有照射水平下都保持了全能，最高可达400rad。由于F-1杂交种ES细胞亲本微卫星间的多态性对于确定诱导缺失的大小具有重要意义，因此我们测定了染色体(Chr)17上38个微卫星座位的129/SVJ和129/Sv(Jae)等位基因的大小。这揭示了CHR 17在129和C57BL/6J之间存在较高水平的多态性。辐射耐受性、亲本株之间的高度多态以及广泛使用的C57BL/6J株组分的存在使v6.3 ES细胞成为具有吸引力的产生辐射诱发染色体缺失的细胞系。
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.