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[人工酶在基因组工程中的应用及潜力]

[Application and potential of genome engineering by artificial enzymes].

作者信息

Nomura Wataru

机构信息

Institute of Biomaterials and Bioengineering, Tokyo Medical and Dental University.

出版信息

Yakugaku Zasshi. 2015;135(3):405-14. doi: 10.1248/yakushi.14-00240-5.

Abstract

Artificial zinc finger proteins (ZFPs) consist of Cys2-His2-type modules composed of approximately 30 amino acids that adopt a ββα structure and coordinate a zinc ion. ZFPs recognizing specific DNA target sequences can substitute for the binding domains of various DNA-modifying enzymes to create designer nucleases, recombinases, and methylases with programmable sequence specificity. Enzymatic genome editing and modification can be applied to many fields of basic research and medicine. The recent development of new platforms using transcription activator-like effector (TALE) proteins or the clustered regularly interspaced short palindromic repeats/CRISPR-associated protein (CRISPR/Cas) system has expanded the range of possibilities for genome-editing technologies. These technologies empower investigators with the ability to efficiently knockout or regulate the functions of genes of interest. In this review, we discuss historical advancements in artificial ZFP applications and important issues that may influence the future of genome editing and engineering technologies. The development of artificial ZFPs has greatly increased the feasibility of manipulating endogenous gene functions through transcriptional control and gene modification. Advances in the ZFP, TALE, and CRISPR/Cas platforms have paved the way for the next generation of genome engineering approaches. Perspectives for the future of genome engineering are also discussed, including applications of targeting specific genomic alleles and studies in synthetic biology.

摘要

人工锌指蛋白(ZFPs)由约30个氨基酸组成的Cys2-His2型模块构成,这些模块采用ββα结构并配位一个锌离子。识别特定DNA靶序列的ZFPs可替代各种DNA修饰酶的结合结构域,以创建具有可编程序列特异性的定制核酸酶、重组酶和甲基化酶。酶促基因组编辑和修饰可应用于基础研究和医学的许多领域。最近利用转录激活样效应因子(TALE)蛋白或成簇规律间隔短回文重复序列/CRISPR相关蛋白(CRISPR/Cas)系统开发的新平台,扩大了基因组编辑技术的可能性范围。这些技术使研究人员能够有效地敲除或调节感兴趣基因的功能。在本综述中,我们讨论了人工ZFP应用的历史进展以及可能影响基因组编辑和工程技术未来的重要问题。人工ZFP的发展极大地提高了通过转录控制和基因修饰来操纵内源基因功能的可行性。ZFP、TALE和CRISPR/Cas平台的进展为下一代基因组工程方法铺平了道路。我们还讨论了基因组工程的未来前景,包括靶向特定基因组等位基因的应用和合成生物学研究。

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