TALE核酸酶介导的基因组定点修饰技术  

Genome Targeting Modification Technology Based on TALE Nuclease Engineering

作　　者：王昕[1] 张志强[1] 张智英[1] 

机构地区：[1]西北农林科技大学动物科技学院,陕西杨凌712100

出　　处：《中国生物化学与分子生物学报》2012年第3期211-216,共6页Chinese Journal of Biochemistry and Molecular Biology

基　　金：国家重点基础研究发展计划(973计划;No.2011CBA01002)~~

摘　　要：TALE核酸酶(TALE nucleases,TALENs)是继锌指核酸酶(zinc-finger nucleases,ZFNs)技术以来的另一种能够对基因组进行高效定点修饰的新技术.TALENs技术是基于植物病原体黄单胞菌(Xanthomonas)分泌的一种转录激活子样效应因子(transcription activator–like effectors,TALEs)而设计和构建的.TALEs蛋白是由N端分泌信号、中央DNA结合域、1个核定位信号和C端激活域构成;TALE核酸酶是将TALE蛋白中的DNA结合域与FokI核酸内切酶的切割域融合,设计和构建的能在特定位点产生双链断裂的TALENs.研究结果显示,人工构建的TALENs能够对动植物细胞的基因组进行高度特异和高效的修饰.TALENs的活性在酵母、植物和哺乳动物包括人的细胞中已经相继得到验证.本文介绍了TALENs的结构特点、作用原理、构建方法和双链断裂的修复机制及其在动植物细胞基因组定点修饰中的应用,剖析了该技术可能存在的问题,展望了TALENs应用前景.TALE nuclease（TALENs） is another new technology to specifically and effectively modify genome following zinc figure nuclease technology established.TALEN technology is based on the discovery of TALEs（transcription activator–like effectors） secreted by plant pathogen Xanthomonas.TALEs have specific structural features,including the N-terminal secretion signal,a DNA binding domain,a nuclease localization signal and acidic activation domain at the C-terminus of the protein.The DNA binding domain of TALEs fused to the FokI cleavage domain can be used to generate chimeric nuclease（TALENs） that bind to the DNA and create double strand breaks（DSBs）.It has been demonstrated that artificial TALENs can specifically and effectively modify genome.The activities of TALENs have been tested in yeast,plant and mammalian cells including human cells.This review summarizes the structure of TALEs protein and its mechanism of functions and the assembly method of TALENs,and briefly describes the cell repair mechanism mediated by double-strand break,and their application in genome editing.The potential challenges and wide application of TALENs technology in the future are prospected.

关 键 词：TALE核酸酶 基因组 定点修饰 

分 类 号：Q783.1[生物学—分子生物学]