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nCas9 Engineering for Improved Target Interaction Presents an Effective Strategy to Enhance Base Editing

Adv Sci (Weinh). 2024-06; 
Guiquan Zhang, Ziguo Song, Shisheng Huang, Yafeng Wang, Jiayuan Sun, Lu Qiao, Guanglei Li, Yuanyuan Feng, Wei Han, Jin Tang, Yulin Chen, Xingxu Huang, Furui Liu, Xiaolong Wang, Jianghuai Liu
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Synthetic sgRNA and crRNA Service … The sgRNA and GFP mRNA (as positive CTRL to indicate LNP delivery) used were synthesized from GenScript (Nanjing, China). Spacer sequences of sgRNA are listed in Tables S4. … Get A Quote

摘要

Base editors (BEs) are a recent generation of genome editing tools that couple a cytidine or adenosine deaminase activity to a catalytically impaired Cas9 moiety (nCas9) to enable specific base conversions at the targeted genomic loci. Given their strong application potential, BEs are under active developments toward greater levels of efficiency and safety. Here, a previously overlooked nCas9-centric strategy is explored for enhancement of BE. Based on a cytosine BE (CBE), 20 point mutations associated with nCas9-target interaction are tested. Subsequently, from the initial positive X-to-arginine hits, combinatorial modifications are applied to establish further enhanced CBE variants (1.1-1.3). Parallel nCas9 m... More

关键词

T‐cells, base editor, enhanced BE, nCas9 engineering, non‐targeted strand