HIGH-FREQUENCY TARGETING OF GENES TO SPECIFIC SITES IN THE MAMMALIAN GENOME
HIGH-FREQUENCY TARGETING OF GENES TO SPECIFIC SITES IN THE MAMMALIAN GENOME
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DOI：
10.1016/0092-8674(86)90463-0
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发表时间：
1986-02-14
期刊：
影响因子：
64.5
通讯作者：
CAPECCHI, MR
中科院分区：
文献类型：
--
作者：
THOMAS, KR;FOLGER, KR;CAPECCHI, MR
我们通过将携带不同突变的同一基因的拷贝注入细胞核，纠正了哺乳动物细胞染色体中的缺陷基因。我们确定了整合基因的数量、排列和染色体位置以及注射分子的数量如何影响基因靶向频率。新引入的DNA与其染色体同源物之间的分离发生的频率为1/103接受DNA的细胞。校正事件由双相互重组或基因转换介导。这导致基因组中的序列被引入的DNA的序列取代，或者在单独的实验中，引入的DNA中的序列被染色体序列取代。点突变和缺失突变均得到纠正;然而，相应序列携带的突变性质影响整合或注入序列是否得到纠正。
We corrected a defective gene residing in the chromosome of a mammalian cell by injecting into the ucleus copies of the same gene carrying a different mutation. We determined how the number, the arrangement, and the chromosomal position of the integrated gene, as well as the number of injected molecules influence the gene-targeting frequency. Recombination between the newly introduced DNA and its chromosomal homolog occurred at a frequency of 1 in 103 cells receiving DNA. Correction events were mediated by either double reciprocal recombination or gene conversion. This resulted in sequences in the genome being replaced by sequences of the introduced DNA or, in separate experiments, sequences in the incoming DNA being replaced by chromosomal sequences. Both point mutations and deletion mutations were corrected; however, the nature of the mutation carried by the respective sequence influenced whether the integrated or injected sequence was corrected.