Increasing the targeting scope and efficiency of base editing with Proxy-BE strategy

FEBS Letters

article

Yin Liu¹  Guanglei Li²  Guang Yang²  Huifeng Gu¹  Shisheng Huang²  Wenxia Yu²  Guizhen Qin¹  Xinyi Liu³  Fuling Zhou⁴  Xingxu Huang²  Yongchang Wei¹ 

[1] Department of Radiation and Medical Oncology, Zhongnan Hospital, Wuhan University;School of Life Science and Technology, ShanghaiTech University;Department of Gastroenterology, Second Clinical Medical College, Jinan University, Shenzhen People's Hospital;Department of Hematology, Zhongnan Hospital of Wuhan University

关键词: base editing;    Cas9;    CRISPR;    dCpf1-BE;    gene editing;    SaKKHBE3;   

DOI  :  10.1002/1873-3468.13719

来源: John Wiley & Sons Ltd.

【 摘 要 】

Base editors (BEs) are widely used in precise gene editing due to their simplicity and versatility. However, their efficiencies are hindered by various obstacles. Considering the chromatin microenvironment as a possible obstacle, here, we demonstrate a further development of the proxy-clustered regularly interspaced short palindromic repeats strategy, termed Proxy-BE, to increase gene editing efficiency. Specifically, a nuclease-dead Cas9 (dCas9) was bound to the sequence about 20–30 base pair away from the target site, potentially improving access to the DNA and, thus, providing a better editing microenvironment for base editors. Our findings confirm that nuclease-dead Streptococcus pyogenes Cas9 can assist the base editors SaKKH-BE3 and dCpf1-BE to double their canonical base editing efficiency. This work provides a new approach to enhance base editing, extending its scope for biological research and gene therapy.

【 授权许可】

Unknown