| CRISPR-array-mediated imaging of non-repetitive and multiplex genomic loci in living cells | |
| 論文作者 | Yang, LZ; Min, YH; Liu, YX; Gao, BQ; Liu, XQ; Huang, YK; Wang, HF; Yang, L; Liu, ZJ; Chen, LL |
| 期刊/會議名稱 | NATURE METHODS |
| 論文年度 | 2024 |
| 論文類別 | |
| 摘要 | 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. This work introduces CRISPRdelight, a dCas12a-based DNA-imaging tool that facilitates the imaging of non-repetitive loci and the tracking of DNA dynamics. |
| 影響因子 | 36.1 |
官方微信
