Key Laboratory of Rice Genetics & Breeding, Institute of Rice Research, Anhui Academy of Agricultural Science, Hefei, China.
Nat Plants. 2020 Mar;6(3):197-201. doi: 10.1038/s41477-020-0603-9. Epub 2020 Feb 24.
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.
化脓链球菌 Cas9（SpCas9）-NG 识别 NGN 间隔基序邻近基序，并扩展了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，而且还能有效地在转基因植物中的转移 DNA 中自我靶向单指导 RNA 序列，通过产生新的单指导 RNA 潜在地增加脱靶风险。我们进一步表明，通过使用从 GCCCC 序列开始的修饰单指导 RNA 支架，可以大大减轻 SpCas9-NG 的自我靶向效应。
