Production of chimaeric mice containing embryonic stem (ES) cells carrying a homoeobox Hox 1.1 allele mutated by homologous recombination
Production of chimaeric mice containing embryonic stem (ES) cells carrying a homoeobox Hox 1.1 allele mutated by homologous recombination
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DOI：
10.1038/338150a0
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发表时间：
1989-03
期刊：
影响因子：
64.8
通讯作者：
A. Zimmer;P. Gruss
中科院分区：
文献类型：
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作者：
A. Zimmer;P. Gruss
一些与果蝇发育控制基因相关的小鼠基因家族已被克隆并进行了结构分析。基于与果蝇基因的结构相似性及其在小鼠胚胎发生过程中的空间和时间限制性表达模式，有人提出这些哺乳动物基因也参与发育控制1 -4。为了通过遗传学手段阐明同源盒基因的功能，必须产生小鼠突变体。我们开发了一种体内诱变技术，并利用该技术对同源同源盒Hox 1.1基因进行了诱变。体内诱变是通过内源性Hox 1.1等位基因与显微注射的多能胚胎干细胞（ES）中的突变基因之间的同源重组实现的5 -9。通过聚合酶链反应（PCR）10鉴定突变细胞，并使用突变克隆产生嵌合小鼠。因为同源重组事件在形式上是基因转换事件，并且不需要筛选携带突变等位基因的细胞，所以体内诱变允许在不引入任何其他序列的情况下对靶序列进行特异性改变。
SEVERAL mouse gene families related toDrosophiladevelopmental control genes and containing a homoeobox, a paired box or a finger domain, have been cloned and structurally analysed. On the basis of structural similarities to theDrosophilagenes and of their spatially and temporally restricted expression patterns during mouse embryogenesis, it has been proposed that these mammalian genes also are involved in the control of develop-ment1–4. To elucidate the function of homoeobox genes by genetic means, mouse mutants must be generated. We have developed a technique for mutagenesisin vivoand have used it to mutate the homoeoboxHox 1.1gene.In vivomutagenesis was achieved through homologous recombination between an endogenousHox 1.1allele and a microinjected mutated gene in pluripotent embryonic stem (ES) cells5–9. Mutant cells were identified by means of the polymerase chain reaction (PCR)10and mutant clones were used to generate chimaeric mice. Because the homologous recombi-nation event is formally a gene conversion event and no selection is required to screen for cells carrying the mutated allele,in vivomutagenesis allows specific alterations in the target sequence to be made without the introduction of any other sequences.