哺乳动物基因组靶向修饰阳性细胞富集的报告载体系统研究进展

doi:10.16288/j.yczz.15-247

遗传 ›› 2016, Vol. 38 ›› Issue (1): 28-39.doi: 10.16288/j.yczz.15-247

哺乳动物基因组靶向修饰阳性细胞富集的报告载体系统研究进展

白义春¹, 徐坤¹, 魏泽辉¹, 马琤², 张智英¹

1. 西北农林科技大学动物科技学院,杨凌 712100;
2. 陕西省兴平市畜牧局,兴平 713199

收稿日期:2015-06-01 修回日期:2015-10-30 出版日期:2016-01-20 发布日期:2016-01-20
通讯作者: 张智英,博士,教授,研究方向：动物基因编辑。E-mail: zhangzhy@nwsuaf.edu.cn E-mail:yichun1979@126.com
作者简介:白义春,博士研究生,专业方向：动物遗传育种。
基金资助:
国家重点基础研究发展计划(973计划)(编号：2011CBA01002)项目和国家科技重大专项(编号: 2014ZX0801009B)资助

Research progress in developing reporter systems for the enrichment of positive cells with targeted genome modification

Yichun Bai¹, Kun Xu¹, Zehui Wei¹, Zheng Ma², Zhiying Zhang¹

1. College of Animal Science & Technology, Northwest A&
F University, Yangling 712100, China;
2. Xingping Animal Husbandry Bureau of Shaanxi Province,Xingping 713199, China

Received:2015-06-01 Revised:2015-10-30 Online:2016-01-20 Published:2016-01-20
Supported by:
[Supported by the National Basic Research Program of China (973 Program) (No; 2011CBA01002) and the National Science and Technology Major Project of China (No; 2014ZX0801009B)]

摘要/Abstract

摘要： 基因组靶向修饰技术对基因功能研究、基因治疗以及转基因育种研究都具有重要的意义和价值。近年来发展起来的人工核酸酶如ZFNs、TALENs和CRISPR/Cas9等的应用大大提高了基因组靶向修饰的效率。但是由于核酸酶表达载体转染效率、核酸酶表达效率及活性以及基因组被打靶后的修复效率等因素在一定程度上制约着基因组靶向修饰阳性细胞的获得。因此富集和筛选基因组靶向修饰阳性细胞是一个亟待解决的问题。报告载体系统可以间接地反映核酸酶的工作效率并有效富集核酸酶修饰的阳性细胞,进而提高基因组靶向修饰阳性细胞的富集和筛选效率。本文主要针对由非同源末端连接(Non-homologous end joining,NHEJ)和单链退火(Single-strand annealing,SSA)两种修复机制分别介导的报告载体系统的原理和应用进行了详细的介绍,以期为以后的相关研究提供借鉴和参考。

关键词: 基因组靶向修饰, 人工核酸酶, 报告载体系统, 阳性细胞富集

Abstract: Targeted genome editing technology plays an important role in studies of gene function, gene therapy and transgenic breeding. Moreover, the efficiency of targeted genome editing is increased dramatically with the application of recently developed artificial nucleases such as ZFNs, TALENs and CRISPR/Cas9. However, obtaining positive cells with targeted genome modification is restricted to some extent by nucleases expression plasmid transfection efficiency, nucleases expression and activity, and repair efficiency after genome editing. Thus, the enrichment and screening of positive cells with targeted genome modification remains a problem that need to be solved. Surrogate reporter systems could be used to reflect the efficiency of nucleases indirectly and enrich genetically modified positive cells effectively, which may increase the efficiency of the enrichment and screening of positive cells with targeted genome modification. In this review, we mainly summarized principles and applications of reporter systems based on NHEJ and SSA repair mechanisms, which may provide references for related studies in future.

Key words: targeted genome modification, engineered nucleases, surrogate reporter system, enrichment of positive cells

白义春, 徐坤, 魏泽辉, 马琤, 张智英. 哺乳动物基因组靶向修饰阳性细胞富集的报告载体系统研究进展[J]. 遗传, 2016, 38(1): 28-39.

Yichun Bai, Kun Xu, Zehui Wei, Zheng Ma, Zhiying Zhang. Research progress in developing reporter systems for the enrichment of positive cells with targeted genome modification[J]. HEREDITAS(Beijing), 2016, 38(1): 28-39.

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哺乳动物基因组靶向修饰阳性细胞富集的报告载体系统研究进展

Research progress in developing reporter systems for the enrichment of positive cells with targeted genome modification

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