摘要
The advent of CRISPR-Cas has revolutionized precise gene editing.While pioneering CRISPR nucleases like Cas9 and Cas12 generate targeted DNA double-strand breaks(DSB)for knockout or homology-directed repair,next generation CRISPR technologies enable gene editing without DNA DSB.Base editors directly convert bases,prime editors make diverse alterations,and dead Cas-regulator fusions allow nuanced control of gene expression,avoiding potentially risks like translocations.Meanwhile,the discovery of diminutive Cas12 orthologs and Obligate Mobile Element-Guided Activity(OMEGA)nucleases has overcome cargo limitations of adeno-associated viral vectors,expanding prospects for in vivo therapeutic delivery.Here,we review the ever-evolving landscape of cutting-edge gene editing tools,focusing on miniature Cas12 orthologs and OMEGA effectors amenable to single AAV packaging.We also summarize CRISPR therapies delivered using AAV vectors,discuss challenges such as efficiency and specificity,and look to the future of this transformative field of in vivo gene editing enabled by AAV vectors delivery.
基金
supported by National Science and Technology Innovation 2030 Major Program (2021ZD0200900) (H.Y.)
the National Natural Science Foundation of China (31925016,82021001) (H.Y.)
Basic Frontier Scientific Research Program of Chinese Academy of Sciences From 0 to 1 original innovation project (ZDBS-LY-SM001) (H.Y.)
Project of Shanghai Municipal Science and Technology Commission (20MC1920400) (H.Y.)
Huida Gene Therapeutics Co.
Ltd
support from Shanghai Center for Brain Science and BrianInspired Technology。
作者简介
Corresponding authors:Xiangfeng Kong,email:kongxiangfeng@simm.ac.cn;Corresponding authors:Hui Yang,email:huiyang@ion.ac.cn。
