High-fidelity gene targeting in embryonic stem cells by using sequence replacement vectors.

作者：K R Thomas, Chu‐Xia Deng, Mario R. Capecchi · 发表于：Molecular and Cellular Biology · 年份：1992 · DOI：10.1128/mcb.12.7.2919 · 被引用次数：139 · 研究领域：Animal Genetics and Reproduction、CRISPR and Genetic Engineering、Pluripotent Stem Cells Research

Mutations were targeted to the Hprt locus in murine embryonic stem cells by using sequence replacement vectors. When the vector was designed such that the mutated sequences were flanked on both sides by several kilobases of DNA homologous to the target locus, replacement of chromosomal sequences with the exogenous DNA occurred with precision. If, on the other hand, the target-homologous DNA on one arm of the vector was reduced to below 1 kb in length, the fidelity of recombination was diminished.