Kong Xiangfeng, Li Tong, Yang Hui
Shanghai Institute of Materia Medica, Chinese Academy of Sciences, Shanghai, 201203, China.
Institute of Neuroscience, Center for Excellence in Brain Science and Intelligence Technology, Chinese Academy of Sciences, Shanghai, 200031, China.
Sci China Life Sci. 2024 Dec;67(12):2540-2553. doi: 10.1007/s11427-023-2608-5. Epub 2024 Sep 27.
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.
CRISPR-Cas的出现彻底改变了精确基因编辑。虽然像Cas9和Cas12这样的开创性CRISPR核酸酶会产生靶向DNA双链断裂（DSB）用于基因敲除或同源定向修复，但新一代CRISPR技术能够在不产生DNA DSB的情况下进行基因编辑。碱基编辑器直接转换碱基，引导编辑器进行多种改变，而无活性的Cas调节因子融合体则允许对基因表达进行细微调控，避免了易位等潜在风险。与此同时，微小Cas12直系同源物和专性移动元件引导活性（OMEGA）核酸酶的发现克服了腺相关病毒载体的货物限制，扩大了体内治疗递送的前景。在这里，我们回顾了前沿基因编辑工具不断演变的情况，重点关注适合单个腺相关病毒包装的微小Cas12直系同源物和OMEGA效应器。我们还总结了使用腺相关病毒载体进行的CRISPR治疗，讨论了效率和特异性等挑战，并展望了由腺相关病毒载体递送实现的体内基因编辑这一变革性领域的未来。
