摘要
真核细胞的基因打靶是基因结构与功能研究的一种非常有价值的技术,也是可应用于基因治疗的具有潜力的工具。有2个限制因素束缚真核细胞基因打靶的发展,即同源重组(HR)率非常低而随机整合率非常高。通过特定基因的过表达或表达干涉,使一些参与DNA重组的蛋白表达水平瞬间改变,可能会增加HR率,降低随机整合率。本文列举了一些与HR相关的候选基因,详细介绍了其中的Rad52上位簇基因,还讨论了打靶载体的设计与修饰、DNA转染方法的有效性等。
Gene targeting in vertebrate cells has proven invaluable in biotechnology, in studies of gene structure and function. It also offers a potential tool for gene-therapeutic applications. Two limitations constrain the current technology: the low rate of homologous recombination in vertebrate cells and the high rate of random (nontargeted) integration of the vector DNA. The possible ways to overcome these limitations within the framework of our present understanding of recombination mechanisms and machinery were reviewed. Several studies suggest that transient alteration of the levels of recombination proteins, by overexpression or interference with expression, may be able increase homologous recombination or decrease random integration, and we present a list of candidate genes. The potentially beneficial modifications to the vector DNA and the effects of methods of DNA delivery on targeting efficiency were discussed.
出处
《生物技术通讯》
CAS
2006年第1期88-91,共4页
Letters in Biotechnology
基金
国家高技术研究发展计划重大专项(2002AA206621)
关键词
同源重组
基因打靶
双链断裂修复
非同源末端结合
homologous recombination
gene targeting
double-strand break repair
nonhomologous end joining