Mutanted OsCOL9 Based on CRISPR/Cas9 Technology in Rice

doi:10.7668/hbnxb.2017.04.007

Abstract

Abstract: The CRISPR/Cas9 system can directly to edit endogenous genomic loci in plant,provide a new opportunities for crop breeding.To explore the editing efficiency of the system on rice endogenous gene and obtain meaningful mutants,using the susceptible rice variety Lijiang and the disease resistance gene OsCOL9 as the target gene.The full-length CDS of OsCOL9 was 1 266 bp long and encoding a 422 aa protein with a molecular weight of about 45.6 kDa,and the N-terminus contained the B-box domain,and the C-terminus contains the CCT (CONSTANS,CONSTANS-Like,TOC1) domain.In this study,four 20 nt guide RNAs (gRNAs) which were targeted to the initiation region CDS and terminal exon of OsCOL9 were designed and transcribed from the U3,U6a,U6b and U6c promoters,respectively. The sequences near the editing site were analyzed by extracting genomic DNA from T1 transgenic plants.The number of target base deletions was 59 bp and the number of mutations was up to 11 times.In this study,we get better gene editing results.However,there have been more serious off-target effects in this experiment.Therefore,we focus on off-target effects in the discussion section of this paper. Owing largely to its flexibility,efficiency and no insertion of exogenous gene,it has important practical significance to combine biotechn ology and traditional breeding.

Key words: Rice, Genome editing, CRISPR/Cas9, OsCOL9

CLC Number:

Q78
S572.03

LIU Wei, LIU Hao, DONG Shuangyu, GU Fengwei, CHEN Zhiqiang, WANG Jiafeng, WANG Hui. Mutanted OsCOL9 Based on CRISPR/Cas9 Technology in Rice[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2017, 32(4): 42-48. doi: 10.7668/hbnxb.2017.04.007.

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