期刊
CROP JOURNAL
卷 8, 期 3, 页码 403-407出版社
KEAI PUBLISHING LTD
DOI: 10.1016/j.cj.2019.06.007
关键词
crRNA; CRISPR-Cas12a; tRNA-crRNA array; Genome editing; Editing efficiency
资金
- National Key Research and Development Program of China [2016YFD0101800]
- Agricultural Science and Technology Innovation Program of Chinese Academy of Agricultural Sciences
- National GMO New Variety Breeding Program of China [2016ZX08011-001]
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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