Use of gene targeting to study recombination in mammalian cell DNA repair mutants.
Use of gene targeting to study recombination in mammalian cell DNA repair mutants.
复制标题
使用基因打靶研究哺乳动物细胞 DNA 修复突变体的重组。
DOI：
10.1007/978-1-61779-998-3_31
复制
发表时间：
2012
期刊：
影响因子：
--
通讯作者：
Nairn,RodneyS
中科院分区：
文献类型：
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作者：
Rahn,JenniferJ;Adair,GeraldM;Nairn,RodneyS
基因组 DNA 修饰策略的发展极大地促进了基因功能的研究。基因打靶是最成功应用的技术之一，用于检查特定基因在从酵母到哺乳动物的各种模型系统中的作用。我们实验室率先使用中国仓鼠卵巢（CHO）细胞培养模型系统来研究半合子CHOAPRT位点的DNA修复和重组途径。通过使用简单有效的基因打靶方法，我们已经产生了许多DNA修复缺陷的细胞系，这些细胞系已用于靶向重组实验，以研究体细胞哺乳动物细胞的重组修复途径。这些方法可以很容易地定制，以产生各种缺乏特定目标基因的细胞系，并可用于研究其他 DNA 修复蛋白在哺乳动物重组修复途径中的作用。
The study of gene function has been greatly facilitated by the development of strategies to modify genomic DNA. Gene targeting is one of the most successfully applied techniques used to examine the roles of specific genes in a wide variety of model systems from yeast to mammals. Our laboratory has pioneered the use of the Chinese hamster ovary (CHO) cell culture model system to study pathways of DNA repair and recombination at the hemizygous CHOAPRTlocus. By using a simple and effective gene targeting method, we have generated a number of DNA repair-deficient cell lines that have been used in targeted recombination experiments to investigate pathways of recombinational repair in somatic mammalian cells. These methods can be readily customized to generate a variety of cell lines deficient in specific genes of interest and can be applied to study the roles of other DNA repair proteins in pathways of mammalian recombinational repair.