Application of the technology of CRISPR/Cas9 edit rice gene

doi:10.3969/j.issn.1004-1524.2017.05.01

›› 2017, Vol. 29 ›› Issue (5): 685-693.DOI: 10.3969/j.issn.1004-1524.2017.05.01

• Crop Science • Next Articles

Application of the technology of CRISPR/Cas9 edit rice gene

YUAN Wenxia^{1, 2}, WANG Xuming², LI Dongyue^{2, 3}, ZHOU Jie², YAN Chengqi^{1, 2, *}, CHEN Jianping^{1, 2, *}

1.College of Chemistry and Life Sciences, Zhejiang Normal University, Jinhua 321004, China;
2.State Key Laboratory Breeding Base for Zhejiang Sustainable Pest and Disease Control, MOA Key Laboratory for Plant Protection and Biotechnology, Zhejiang Provincial Key Laboratory of Virology, Institute of Virology and Biotechnology, Zhejiang Academy of Agricultural Sciences, Hangzhou 310021, China;
3.College of Plant Protection, Yunnan Agricultural University, Kunming 650201, China

Received:2017-03-09 Online:2017-05-20 Published:2017-06-06

Abstract

Abstract: In this study, the CRISPR/Cas9 vector system was optimized in order to construct gene editing constructors in two steps. With optimized CRISPR/Cas9 system, three genome editing constructors targeting different sites of D3 gene in rice (Oryza sativa L. spp. Japonica cv. Nipponbare) were generated, and the transgenic plants were successfully produced by Agrobacterium-mediated transformation. The sequencing results of T₀ transgenic plants showed that the mutation rate of target site DDRC1 was 35.48%, and five mutation types were detected; the mutation rate of target site DDRC2 was 25.00%, and also with five mutation types; the mutation rate of target site DDRC3 was 20.00%, and only one mutation type was detected. The phenotype, height and tiller number, together with the genotypes of homozygous mutants were investigated, and the gene editing efficiency of the optimized CRISPR/Cas9 system along with the position effect and mutation patterns were also analyzed and discussed. In summary, the findings in this research provided important information for further developing and utilizing of rice germplasm on variety agronomic trait with the optimized CRISPR/Cas9 system.

Key words: rice, CRISPR/Cas9, D3 gene, gene editing

CLC Number:

S511

YUAN Wenxia, WANG Xuming, LI Dongyue, ZHOU Jie, YAN Chengqi, CHEN Jianping. Application of the technology of CRISPR/Cas9 edit rice gene[J]. , 2017, 29(5): 685-693.

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Application of the technology of CRISPR/Cas9 edit rice gene

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