Xiao An, Hu Ying-Ying, Wang Wei-Ye, Yang Zhi-Peng, Wang Zhan-Xiang, Huang Peng, Tong Xiang-Jun, Zhang Bo, Lin Shuo
Key Laboratory of Cell Proliferation and Differentiation of Ministry of Education, College of Life Sciences, Peking University, Beijing 100871, China.
Yi Chuan. 2011 Jul;33(7):665-83. doi: 10.3724/sp.j.1005.2011.00665.
Zinc finger nuclease (ZFN) is an artificially engineered hybrid protein that contains a zinc finger protein (ZFP) domain and a Fok I endonuclease cleavage domain. It has recently emerged as a powerful molecular tool for targeted genome modifications. ZFNs recognize and bind to specific DNA sequences to generate a double-strand break (DSB) by its nuclease activity. Based on this finding, various genetic methods, including gene targeting (gene disruption), gene addition, gene correction etc., are being designed to manipulate the genomes of different species at specific loci. One particular advantage of this new technique is its broad applications, which can be employed to generate desirable inheritable mutations both at the organismal level and at the cellular level. Here, we review the recent progress and prospects of ZFN technology. This article focused on the mechanism of how it works, currently available target assessment, ZFP library construction and screening methods, target modification strategies, as well as a collection of specie and genes that have been successfully modified by ZFN. This review will provide a useful reference for researchers who are interested in applying this new technique in their studies.
锌指核酸酶(ZFN)是一种人工构建的杂合蛋白,它包含一个锌指蛋白(ZFP)结构域和一个Fok I核酸内切酶切割结构域。最近,它已成为一种用于靶向基因组修饰的强大分子工具。ZFN通过其核酸酶活性识别并结合特定的DNA序列,以产生双链断裂(DSB)。基于这一发现,人们正在设计各种遗传方法,包括基因靶向(基因破坏)、基因添加、基因校正等,以在特定位点操纵不同物种的基因组。这项新技术的一个特别优点是其广泛的应用,可用于在生物体水平和细胞水平产生理想的可遗传突变。在此,我们综述了ZFN技术的最新进展和前景。本文重点关注其作用机制、当前可用的靶点评估、ZFP文库构建和筛选方法、靶点修饰策略,以及已被ZFN成功修饰的一系列物种和基因。本综述将为有兴趣在其研究中应用这项新技术的研究人员提供有用的参考。