Improving the efficiency of the CRISPR-Cas12a system with tRNA-crRNA arrays

文献类型： 外文期刊

第一作者： Hu, Xixun

作者： Hu, Xixun;Wang, Kejian;Meng, Xiangbing;Li, Jiayang;Yu, Hong;Meng, Xiangbing;Li, Jiayang;Yu, Hong;Li, Jiayang

作者机构：

关键词： crRNA; CRISPR-Cas12a; tRNA-crRNA array; Genome editing; Editing efficiency

期刊名称：CROP JOURNAL （ 影响因子：4.407； 五年影响因子：5.687 ）

ISSN： 2095-5421

年卷期： 2020 年 8 卷 3 期

页码：

收录情况： SCI

摘要： CRISPR-Cas12a offers a convenient tool for multiplex genome editing in rice. However, the CRISPR-Cas12a system displays variable editing efficiency across genomic loci, with marked influence by CRISPR RNAs (crRNAs). To improve the efficiency of the CRISPR-Cas12a system for multiplex genome editing, we identified various architectures and expression strategies for crRNAs. Transformation of binary vectors loaded with engineered CRISPR-Cas12a systems into rice calli and subsequent sequencing revealed that a modified tRNA-crRNA array not only efficiently achieved rice multiplex genome editing, but also successfully edited target sites that were not edited by the crRNA array. This improvement contributes to the application of the CRISPR-Cas12a system in plant genome editing, especially for genomic loci that have hitherto been difficult to edit. (C) 2019 Crop Science Society of China and Institute of Crop Science, CAAS. Production and hosting by Elsevier B.V. on behalf of KeAi Communications Co., Ltd.

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