利用CRISPR-脱氨酶在番茄中进行靶向碱基编辑
Targeted Base Editing with CRISPR-Deaminase in Tomato.
作者信息
Shimatani Zenpei, Ariizumi Tohru, Fujikura Ushio, Kondo Akihiko, Ezura Hiroshi, Nishida Keiji
机构信息
Graduate School of Science, Technology and Innovation, Kobe University, Kobe, Hyogo, Japan.
Faculty of Life and Environmental Sciences, Gene Research Center, University of Tsukuba, Tsukuba, Ibaraki, Japan.
出版信息
Methods Mol Biol. 2019;1917:297-307. doi: 10.1007/978-1-4939-8991-1_22.
The Target-AID system, consisting of a complex of cytidine deaminase and deficient CRISPR/Cas9, enables highly specific genomic nucleotide substitutions without the need for template DNA. The Cas9-fused cytidine deaminase is guided by sgRNAs and catalyzes the conversion of cytosine to uracil. The resulting U-G DNA mismatches trigger nucleotide substitutions (C to T or G to A) through DNA replication and repair pathways. Target-AID also retains the benefits of conventional CRISPR/Cas9 including robustness in various organisms, high targeting efficiency, and multiplex simultaneous gene editing. Our research group recently developed plant-optimized Target-AID system and demonstrated targeted base editing in tomato and rice. In this chapter, we introduce methods for Target-AID application in tomato.
靶标-腺嘌呤脱氨酶（Target-AID）系统由胞苷脱氨酶复合物和缺陷型CRISPR/Cas9组成，无需模板DNA即可实现高度特异性的基因组核苷酸替换。与Cas9融合的胞苷脱氨酶由sgRNA引导，并催化胞嘧啶转化为尿嘧啶。由此产生的U-G DNA错配通过DNA复制和修复途径触发核苷酸替换（C到T或G到A）。靶标-腺嘌呤脱氨酶（Target-AID）系统还保留了传统CRISPR/Cas9的优点，包括在各种生物体中的稳健性、高靶向效率和多重同时基因编辑。我们的研究小组最近开发了植物优化的靶标-腺嘌呤脱氨酶（Target-AID）系统，并在番茄和水稻中证明了靶向碱基编辑。在本章中，我们介绍了靶标-腺嘌呤脱氨酶（Target-AID）系统在番茄中的应用方法。
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