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

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SpCas9-NG self-targets the sgRNA sequence in plant genome editing
Nature Plants ( IF 15.8 ) Pub Date : 2020-02-24 , DOI: 10.1038/s41477-020-0603-9
Ruiying Qin ₁ , Juan Li ₁ , Xiaoshuang Liu ₁ , Rongfang Xu ₁ , Jianbo Yang ₁ , Pengcheng Wei ₁

Affiliation

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.

中文翻译：

SpCas9-NG 在植物基因组编辑中自靶向 sgRNA 序列

化脓性链球菌Cas9 (SpCas9)-NG 可识别 NGN 原型间隔区相邻基序并扩展基因组编辑工具的范围。在这项研究中，我们发现SpCas9-NG不仅靶向基因组，而且还有效地自靶向转基因植物中转移DNA中的单向导RNA序列，通过生成新的单向导RNA可能增加脱靶风险。我们进一步表明，通过使用以 GCCCC 序列开始的改良单引导 RNA 支架，可以大大减轻 SpCas9-NG 的自靶向效应。

更新日期：2020-02-24

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