Efficient Cre-IoxP-induced mitotic recombination in mouse embryonic stem cells
Efficient Cre-IoxP-induced mitotic recombination in mouse embryonic stem cells
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DOI：
10.1038/ng788
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发表时间：
2002-01-01
期刊：
影响因子：
30.8
通讯作者：
Copeland, NG
中科院分区：
文献类型：
--
作者：
Liu, PT;Jenkins, NA;Copeland, NG
FLP/FRT诱导的有丝分裂重组为在果蝇中创造遗传嵌合体和识别杂合个体中隐性基因的功能提供了强有力的方法。在此，我们发现，在小鼠胚胎干细胞（ES）中，通过Cre/loxP技术，在Cre瞬时表达后，有丝分裂重组可以重复诱导，频率范围为4.2 x 10（-5）（Snrpn）至7.0 x 10（-3）（D 7 Mit 178）（单个等位基因loxP位点）和5.0 x 10（-2）（D 7 Mit 178）（多个等位基因lox位点）。值得注意的是，大部分重组发生在G2中，随后是X分离，其中重组染色单体在有丝分裂期间彼此分离。X分离对遗传镶嵌分析是有用的，因为它从杂合母体中产生纯合突变子细胞的克隆。我们的研究证实了果蝇研究中的预测（1），即X染色体分离并不局限于有丝分裂配对较强的生物体，因为导致X染色体分离的力（姐妹染色单体凝聚力）是所有真核生物有丝分裂的基本特征。我们的研究还表明，在小鼠中进行遗传嵌合体分析是可行的，至少在某些染色体区域是可行的。
FLP/FRT-induced mitotic recombination provides a powerful method for creating genetic mosaics in Drosophila and for discerning the function of recessive genes in a heterozygous individual. Here we show that mitotic recombination can be reproducibly induced in mouse embryonic stem (ES) cells, by Cre/loxP technology, at frequencies ranging from 4.2 x 10(-5) (Snrpn) to 7.0 x 10(-3) (D7Mit178) for single allelic loxP sites, and to 5.0 x 10(-2) (D7Mit178) for multiple allelic lox sites, after transient Cre expression. Notably, much of the recombination occurs in G2 and is followed by X segregation, where the recombinant chromatids segregate away from each other during mitosis. It is X segregation that is useful for genetic mosaic analysis because it produces clones of homozygous mutant daughter cells from heterozygous mothers. Our studies confirm the predictions made from studies in Drosophila(1) that suggest that X segregation will not be limited to organisms with strong mitotic pairing, because the forces (sister-chromatid cohesion) responsible for X segregation are an elemental feature of mitosis in all eukaryotes. Our studies also show that genetic mosaic analysis in mice is feasible, at least for certain chromosomal regions.