脱氧核酶
原位
清脆的
生物正交化学
Cas9
活体细胞成像
化学
细胞核
纳米技术
生物物理学
DNA
细胞
材料科学
生物化学
生物
组合化学
点击化学
有机化学
基因
作者
Ran Liu,Difei Jiang,Yangfang Yun,Zhe Feng,Fenfen Zheng,Yu Xiang,Huanhuan Fan,Jingjing Zhang
标识
DOI:10.1002/anie.202315536
摘要
Abstract DNAzyme‐based fluorescent probes for imaging metal ions in living cells have received much attention recently. However, employing in situ metal ions imaging within subcellular organelles, such as nucleus, remains a significant challenge. We developed a three‐stranded DNAzyme probe (TSDP) that contained a 20‐base‐pair (20‐bp) recognition site of a CRISPR/Cas9, which blocks the DNAzyme activity. When Cas9, with its specialized nuclear localization function, forms an active complex with sgRNA within the cell nucleus, it cleaves the TSDP at the recognition site, resulting in the in situ formation of catalytic DNAzyme structure. With this design, the CRISPR/Cas9‐inducible imaging of nuclear Zn 2+ is demonstrated in living cells. Moreover, the superiority of CRISPR‐DNAzyme for spatiotemporal control imaging was demonstrated by integrating it with photoactivation strategy and Boolean logic gate for dynamic monitoring nuclear Zn 2+ in both HeLa cells and mice. Collectively, this conceptual design expands the DNAzyme toolbox for visualizing nuclear metal ions and thus provides new analytical methods for nuclear metal‐associated biology.
科研通智能强力驱动
Strongly Powered by AbleSci AI