A genetic screen identifies novel non-compatible IoxP sites
A genetic screen identifies novel non-compatible IoxP sites
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DOI：
10.1093/nar/gkf421
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发表时间：
2002-07-15
影响因子：
14.9
通讯作者：
Leinwand, L
中科院分区：
文献类型：
--
作者：
Langer, SJ;Ghafoori, AP;Leinwand, L
Cre/lox系统进行精确基因组修饰的能力是一项巨大的成就。然而，顺式连接的异源特异性lox位点之间的重组限制了Cre介导的DNA交换系统的使用，其中可以应用遗传选择。为了避免这个问题，我们进行了遗传筛选，旨在确定新的突变间隔区的lox网站展示增强不相容性与典型的loxP网站。回收的突变位点之一似乎在组成型表达Cre重组酶的HEK 293细胞中完全稳定，并支持细菌和哺乳动物细胞培养物中的重组酶介导的盒交换（RMCE）。通过防止不期望的重组，这些新的lox位点可以提高体内基因转移的效率。
The ability of the Cre/lox system to make precise genomic modifications is a tremendous accomplishment. However, recombination between cis-linked heterospecific lox sites limits the use of Cre-mediated exchange of DNA to systems where genetic selection can be applied. To circumvent this problem we carried out a genetic screen designed to identify novel mutant spacer-containing lox sites displaying enhanced incompatibility with the canonical loxP site. One of the mutant sites recovered appears to be completely stable in HEK293 cells constitutively expressing Cre recombinase and supports recombinase-mediated cassette exchange (RMCE) in bacteria and mammalian cell culture. By preventing undesirable recombination, these novel lox sites could improve the efficiency of in vivo gene transfer.