Rationally designed Campylobacter jejuni Cas9 enables efficient gene activation and base editing

作者：Yuxi Chen, Rui Kang, Yuanling Jiang, Qi Zheng, Yue Yang, Jiaqi Liu, Guanglan Wu, Weijun Zhao, Zhan Li, Chengxiang Peng, Pengfei Zhang, Peng Fei, Qianyi Liu, Sihui Hu, Xiao Luo, Guifang Wu, Kaixin Cui, Junjiu Huang, Yongming Wang, Zhou Songyang, Puping Liang · 发表于：Molecular Therapy — Nucleic Acids · 年份：2024 · DOI：10.1016/j.omtn.2024.102366 · 被引用次数：8 · 研究领域：CRISPR and Genetic Engineering、Virus-based gene therapy research、Animal Genetics and Reproduction

is particularly appealing for AAV delivery. However, the editing efficiency of CjCas9 limits its applications. In this study, we used structure-guided protein engineering to improve the editing efficiency of CjCas9. Subsequently, we developed a miniature transcriptional activator (LDE-CjCas9-VPR) and base editors engineered from CjCas9 (LDE-CjABE and LDE-CjCBE). LDE-CjABE effectively induced genome editing in human and mouse cells. Through AAV delivery, LDE-CjABE enhanced the on-target editing efficiency, and off-target editing was not detected in the mouse retina. Therefore, the compact size and high editing efficiency of LDE-CjCas9 broadens the target scope of transcription activation and base editing toolsets for therapeutic applications.