CRISPR/Cas系统在活细胞染色体成像中的应用

Application of CRISPR/Cas9 System in Living Cell Imaging

CRISPR/Cas(Clustered Regularly Interspaced Short Palindromic Repeats/CRISPR-associated Proteins)是细菌抵抗外来入侵的一种自适应免疫机制,利用CRISPR/Cas9系统可实现双链DNA的剪切并诱导宿主细胞DNA修复机制,从而达到靶向编辑基因的目的。核酸酶失活的Cas9(Nuclease-deactivated Cas9,d Cas9)耦联效应分子可以调控靶标结合位点附近基因的表达、表观遗传修饰及特异染色体区域标记。目前已开发出多种CRISPR/Cas9系统,可对活细胞中重复或低重复序列基因位点进行实时多位点同步成像,广泛应用于动物和植物细胞中。基于CRISPR/Cas系统的活细胞染色体成像技术为研究活细胞染色体动力学和三维染色体结构提供了全新角度。本研究针对CRISPR/Cas系统的生源机制及其在活细胞成像的应用和发展现状进行概述,以期为该领域的相关研究提供参考。

CRISPR/Cas is a bacterial adaptive immune system. By cleaving specific strands of DNA and inducing DNA repair mechanism in host cells through CRISPR/Cas9 system, genome editing can be achieved. Nuclease-deactivated Cas9(dCas9) can regulate gene expression, epigenetic modification, and label specific chromosomal regions via coupling effectors. Diverse CRISPR/Cas9 systems can target either the repetitive or low repetitive chromosome loci for live cell visualization, which is applicable to both animal and plant cells. Live cell chromosome imaging based on CRISPR/Cas system provides a new perspective for studying chromosome dynamics and three-dimensional genome structure. This review summarized the recent progress on the study of the molecular mechanism of CRISPR/Cas system and its application in living cell imaging.