J Integr Plant Biol ›› 2021, Vol. 63 ›› Issue (9): 1606-1610.DOI: 10.1111/jipb.13166

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  • 收稿日期:2021-08-07 接受日期:2021-08-23 出版日期:2021-09-01 发布日期:2021-09-22

CRISPR/Sc++-mediated genome editing in rice

Guigen Ma1,2†*, Yongjie Kuang1,2†, Zhenwan Lu1,2, Xueqi Li1,2, Ziyan Xu1,2, Bin Ren1,2, Xueping Zhou1,3 and Huanbin Zhou1,2*   

  1. 1 State Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Beijing 100193, China
    2 Scientific Observing and Experimental Station of Crop Pests in Guilin, Ministry of Agriculture and Rural Affairs, Guilin 541399, China
    3 State Key Laboratory of Rice Biology, Institute of Biotechnology, Zhejiang University, Hangzhou 310058, China

    †These authors contributed equally to this work.
    *Correspondence: Huanbin Zhou (hbzhou@ippcaas.cn)
  • Received:2021-08-07 Accepted:2021-08-23 Online:2021-09-01 Published:2021-09-22

Abstract: Streptococcus canis Cas9 (ScCas9) is an RNA-guided endonuclease with NNG protospacer adjacent motif (PAM) specificity whose genome-editing activity in rice is locus-dependent. Here we investigated the performance of a ScCas9 variant named Sc++ at different NNG PAM sites in the rice genome; Sc++ harbors a T1227K mutation and the substitution of a positively charged loop (residues 367–376). Sc++ nuclease achieved broader genome editing compared to the original ScCas9, and its nickase improved targeted base editing in transgenic rice plants. Using the high-efficiency adenine base editor rBE73b, we generated many new OsGS1 alleles suitable for screening of rice germplasm for potential herbicide resistance in the future. The CRISPR/Sc++ system expands the genome-editing toolkit for rice.

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