中国科学院长春应用化学研究所曲晓刚团队利用CRISPR引导的G-四联体结合蛋白和配体靶向特定DNA G-四联体(G4)。2024年7月3日出版的《自然-细胞生物学》发表了这项成果。
研究人员将G4稳定蛋白/配体与CRISPR结合使用,选择性地调控特定基因组位点内的单个或多个目标G4折叠。研究证明,将核蛋白与催化不活跃的Cas9融合,可以特异性地稳定癌基因MYC和肌肉相关基因Itga7启动子中的G4s以及端粒G4s,从而分别诱导细胞增殖停滞、抑制成肌细胞分化和细胞衰老。
此外,CRISPR还能选择性调控G4与化合物吡啶二甲酰胺和哒哒司汀的结合。与传统的G4配体相比,CRISPR引导的生物素连接的吡啶二甲酰胺能更精确地研究新G4的生物功能。该研究提供的实验结果将有助于加深对G4功能和治疗效果的理解。
据悉,尽管DNA G-四联体在健康和疾病中的重要性已得到证实,但目前仍缺乏可随时操纵特定G4折叠以进行功能分析和治疗的技术。
附:英文原文
Title: Targeting specific DNA G-quadruplexes with CRISPR-guided G-quadruplex-binding proteins and ligands
Author: Qin, Geng, Liu, Zhenqi, Yang, Jie, Liao, Xiaofeng, Zhao, Chuanqi, Ren, Jinsong, Qu, Xiaogang
Issue&Volume: 2024-07-03
Abstract: Despite the demonstrated importance of DNA G-quadruplexes (G4s) in health and disease, technologies to readily manipulate specific G4 folding for functional analysis and therapeutic purposes are lacking. Here we employ G4-stabilizing protein/ligand in conjunction with CRISPR to selectively facilitate single or multiple targeted G4 folding within specific genomic loci. We demonstrate that fusion of nucleolin with a catalytically inactive Cas9 can specifically stabilize G4s in the promoter of oncogene MYC and muscle-associated gene Itga7 as well as telomere G4s, leading to cell proliferation arrest, inhibition of myoblast differentiation and cell senescence, respectively. Furthermore, CRISPR can confer intra-G4 selectivity to G4-binding compounds pyridodicarboxamide and pyridostatin. Compared with traditional G4 ligands, CRISPR-guided biotin-conjugated pyridodicarboxamide enables a more precise investigation into the biological functionality of de novo G4s. Our study provides insights that will enhance understanding of G4 functions and therapeutic interventions.
DOI: 10.1038/s41556-024-01448-1
Source: https://www.nature.com/articles/s41556-024-01448-1
Nature Cell Biology:《自然—细胞生物学》,创刊于1999年。隶属于施普林格·自然出版集团,最新IF:28.213
官方网址:https://www.nature.com/ncb/
投稿链接:https://mts-ncb.nature.com/cgi-bin/main.plex