中国科学院上海生物化学与细胞生物学研究所陈玲玲团队近期取得重要工作进展，他们研究开发了CRISPRdelight，一种由CRISPR阵列介导的活细胞中非重复和多重基因组位点的成像工具。相关研究成果2024年7月4日在线发表于《自然—方法学》杂志上。

据介绍，基因组位点的动态成像是理解基因调控的关键，但对基因组，特别是非重复DNA进行成像的方法是有限的。

研究人员开发了CRISPRelide，这是一种基于核酸内切酶缺陷型CRISPR–Cas12a（dCas12a）的DNA标记系统，具有工程化的CRISPR阵列来跟踪DNA的位置和运动。CRISPRelide能够对不同细胞系中所有检测的12个非重复基因组基因座进行稳定成像。研究人员揭示了CCAT1基因座（chr8q24）在核外周的受限运动，用于抑制表达，并在核内进行主动运动，用于转录。

研究人员发现了HSP基因位点向核斑点的选择性重新定位，包括HSPH1（chr13q12）在应激期间显著的重新定位以提高转录。结合CRISPR–dCas12a和RNA适体，可以用单个阵列对四种类型的卫星DNA基因座进行多重成像，揭示它们与核仁相关结构域的空间接近性。

总之，这一研究表明，CRISPRdelight是一个用户友好且功能强大的系统，用于成像和跟踪基因组动态和调控。

附：英文原文

Title: CRISPR-array-mediated imaging of non-repetitive and multiplex genomic loci in living cells

Author: Yang, Liang-Zhong, Min, Yi-Hui, Liu, Yu-Xin, Gao, Bao-Qing, Liu, Xiao-Qi, Huang, Youkui, Wang, Haifeng, Yang, Li, Liu, Zhe J., Chen, Ling-Ling

Issue&Volume: 2024-07-04

Abstract: Dynamic imaging of genomic loci is key for understanding gene regulation, but methods for imaging genomes, in particular non-repetitive DNAs, are limited. We developed CRISPRdelight, a DNA-labeling system based on endonuclease-deficient CRISPR–Cas12a (dCas12a), with an engineered CRISPR array to track DNA location and motion. CRISPRdelight enables robust imaging of all examined 12 non-repetitive genomic loci in different cell lines. We revealed the confined movement of the CCAT1 locus (chr8q24) at the nuclear periphery for repressed expression and active motion in the interior nucleus for transcription. We uncovered the selective repositioning of HSP gene loci to nuclear speckles, including a remarkable relocation of HSPH1 (chr13q12) for elevated transcription during stresses. Combining CRISPR–dCas12a and RNA aptamers allowed multiplex imaging of four types of satellite DNA loci with a single array, revealing their spatial proximity to the nucleolus-associated domain. CRISPRdelight is a user-friendly and robust system for imaging and tracking genomic dynamics and regulation.

DOI: 10.1038/s41592-024-02333-3

Source: https://www.nature.com/articles/s41592-024-02333-3