AP站点
化学
核酸内切酶
DNA
荧光团
DNA-(无嘌呤或无嘧啶位点)裂解酶
连接器
分子生物学
生物化学
荧光
生物物理学
细胞生物学
生物
物理
操作系统
量子力学
计算机科学
作者
Junjie Li,Wenwen Du,Yining Liu,Fenglin Wang,Li‐Juan Tang,Jian‐Hui Jiang
标识
DOI:10.1021/acs.analchem.2c05504
摘要
Nucleic acids are valuable tools for intracellular biomarker detection and gene regulation. Here we propose a new type of protein (avidin)-scaffolded DNA nanostructure (ADN) for imaging the activity of apurinic/apyrimidinic endonuclease 1 (APE1) in live cells. ADN is designed by assembling an avidin-displayed abasic site containing DNA strands labeled with a fluorophore or a quencher via a complementary linker strand. ADN is nonemissive due to the close proximity of fluorophores and quenchers. APE1-mediated cleavage separates the fluorophores from the quenchers, delivering activated fluorescence. In vitro assays show that ADN is responsive to APE1 with high sensitivity and high specificity. ADN can efficiently enter the cells, and its capability to visualize and detect intracellular APE1 activities is demonstrated in drug-treated cells and different cell lines. The modular and easy preparation of our nanostructures would afford a valuable platform for imaging and detecting APE1 activities in live cells.
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