学术
影响动物细胞同源重组发生与基因打靶效率的分子机制
李兰[1,2] 沈伟[2,3] 邓继先[2] 潘庆杰[1]
[1]莱阳农学院动物科技学院,山东莱阳265200 [2]军事医学科学院生物工程研究所,北京100071 [3]北京大学生命科学学院,北京100871
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。[著者文摘]
文章出处:
《生物技术通讯》-2006年17卷1期 -88-91页
栏目信息:
文献标识码:
A
文章编号:
1009-0002（2006）01-0088-04
Homologous Recombination and Gene Targeting in Vertebrate Cells
LI Lan, SHEN Wei, DENG Ji-xian, PAN Qing-jie（ 1. Department of Animal Science and Technology, Agricultural College of Laiyang, Laiyang 265200; 2. Institute of Biotechnology, Academy of Military Medical Sciences, Beijing 100071; 3. Department of Life Science, Peking University, Beijing 100871, China）
Abstract:
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.[著者文摘]
Key words:
homologous recombination; gene targeting; double-strand break repair; nonhomologous end joining
基金资助:
国家高技术研究发展计划重大专项（2002AA206621）.
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