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SpCas9-NG self-targets the sgRNA sequence in plant genome editing

文献类型： 外文期刊

作者： Qin, Ruiying 1 ; Li, Juan 1 ; Liu, Xiaoshuang 1 ; Xu, Rongfang 1 ; Yang, Jianbo 1 ; Wei, Pengcheng 1 ;

作者机构： 1.Anhui Acad Agr Sci, Inst Rice Res, Key Lab Rice Genet & Breeding, Hefei, Peoples R China

期刊名称：NATURE PLANTS （ 影响因子：15.793； 五年影响因子：17.349 ）

ISSN： 2055-026X

年卷期： 2020 年 6 卷 3 期

页码：

收录情况： SCI

摘要： Streptococcus pyogenes Cas9 (SpCas9)-NG recognizes NGN protospacer adjacent motifs and expands the scope of genome-editing tools. In this study, we found that SpCas9-NG not only targeted the genome but also efficiently self-targeted the single-guide RNA sequence in transfer DNA in transgenic plants, potentially increasing off-target risk by generating new single-guide RNAs. We further showed that the self-target effect of SpCas9-NG could be greatly alleviated by using a modified single-guide RNA scaffold starting with a GCCCC sequence. The widely used SpCas9-NG tool can edit genomic targets, but unfortunately also the guide RNA sequences that are present in the transfer DNA and integrated into the plant genome. The authors propose a strategy to alleviate this self-editing risk.

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