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当前位置: 首页 > 文章 > Improving the efficiency of the CRISPR-Cas12a system with tRNA-crRNA arrays 作物学报(英文版) 2020,8 (3)
Position: Home > Articles > Improving the efficiency of the CRISPR-Cas12a system with tRNA-crRNA arrays The Crop Journal 2020,8 (3)

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

作  者：
Xixun Hu;Xiangbing Meng;Jiayang Li;Kejian Wang;Hong Y
单  位：
State Key Laboratory of Plant Genomics and National Center for Plant Gene Research (Beijing), Institute of Genetics and Developmental Biology, The Innovative Academy of Seed Design, Chinese Academy of Sciences, Beijing 100101, Chin;State Key Laboratory of Rice Biology, China National Rice Research Institute, Chinese Academy of Agricultural Sciences, Hangzhou 310006, Zhejiang, China; University of Chinese Academy of Sciences, Beijing 100049, China;State Key Laboratory of Plant Genomics and National Center for Plant Gene Research (Beijing), Institute of Genetics and Developmental Biology, The Innovative Academy of Seed Design, Chinese Academy of Sciences, Beijing 100101, China
关键词：
crRNA;CRISPR-Cas12a;tRNA-crRNA array;Genome editing;Editing efficienc
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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