A large-scale genome and transcriptome sequencing analysis reveals the mutation landscapes induced by high-activity adenine base editors in plants

文献类型： 外文期刊

第一作者： Li, Shaofang

作者： Li, Shaofang;Liu, Lang;Zhou, Xueping;Zhou, Huanbin;Liu, Lang;Zhou, Huanbin;Liu, Lang;Sun, Wenxian;Zhou, Xueping

作者机构：

关键词： Adenine base editor; TadA variants; Single nucleotide variant; T-DNA insertion; Off-target; Oryza sativa L

期刊名称：GENOME BIOLOGY （ 影响因子：17.906； 五年影响因子：20.367 ）

ISSN： 1474-760X

年卷期： 2022 年 23 卷 1 期

页码：

收录情况： SCI

摘要： Background The high-activity adenine base editors (ABEs), engineered with the recently-developed tRNA adenosine deaminases (TadA8e and TadA9), show robust base editing activity but raise concerns about off-target effects. Results In this study, we perform a comprehensive evaluation of ABE8e- and ABE9-induced DNA and RNA mutations in Oryza sativa. Whole-genome sequencing analysis of plants transformed with four ABEs, including SpCas9n-TadA8e, SpCas9n-TadA9, SpCas9n-NG-TadA8e, and SpCas9n-NG-TadA9, reveal that ABEs harboring TadA9 lead to a higher number of off-target A-to-G (A>G) single-nucleotide variants (SNVs), and that those harboring CRISPR/SpCas9n-NG lead to a higher total number of off-target SNVs in the rice genome. An analysis of the T-DNAs carrying the ABEs indicates that the on-target mutations could be introduced before and/or after T-DNA integration into plant genomes, with more off-target A>G SNVs forming after the ABEs had integrated into the genome. Furthermore, we detect off-target A>G RNA mutations in plants with high expression of ABEs but not in plants with low expression of ABEs. The off-target A>G RNA mutations tend to cluster, while off-target A>G DNA mutations rarely clustered. Conclusion Our findings that Cas proteins, TadA variants, temporal expression of ABEs, and expression levels of ABEs contribute to ABE specificity in rice provide insight into the specificity of ABEs and suggest alternative ways to increase ABE specificity besides engineering TadA variants.

分类号：

  • 相关文献
作者其他论文 更多>>

微信小程序