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

• Main Authors: Yuxi Chen (Author), Rui Kang (Author), Yuanling Jiang (Author), Qi Zheng (Author), Yue Yang (Author), Jiaqi Liu (Author), Guanglan Wu (Author), Weijun Zhao (Author), Zhan Li (Author), Chengxiang Peng (Author), Pengfei Zhang (Author), Fei Peng (Author), Qianyi Liu (Author), Sihui Hu (Author), Xiao Luo (Author), Guifang Wu (Author), Kaixin Cui (Author), Junjiu Huang (Author), Yongming Wang (Author), Zhou Songyang (Author), Puping Liang (Author)
• Format: Book
• Published: Elsevier, 2024-12-01T00:00:00Z.
• Subjects: article
• Online Access: Connect to this object online.

Summary

Compact and adaptable CRISPR-Cas systems enable genome engineering applications in various contexts via high-efficiency delivery. The adeno-associated virus (AAV) is a widely used delivery system. One of the most compact type II-C Cas9 orthologs-CjCas9, derived from Campylobacter jejuni, 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.

Item Description

2162-2531
10.1016/j.omtn.2024.102366