基于DNA功能化金属有机框架材料的荧光探针对瓣状核酸内切酶1的检测和成像研究  

DNA-functionalized Metal-organic Frameworks-based Fluorescent Probe for Detection and Imaging of Flap Endonuclease 1

作　　者：韩庆怡 邵明政 张丁丁 李雨彦 杜佩瑶 张立兵 卢小泉 HAN Qing-Yi;SHAO Ming-Zheng;ZHANG Ding-Ding;LI Yu-Yan;DU Pei-Yao;ZHANG Li-Bing;LU Xiao-Quan(Tianjin Key Laboratory of Molecular Optoelectronic Science,Department of Chemistry,School of Science,Tianjin University,Tianjin 300072,China;Key Laboratory of Bioelectrochemistry&Environmental Analysis of Gansu Province,College of Chemistry&Chemical Engineering,Northwest Normal University,Lanzhou 730070,China)

机构地区：[1]天津大学理学院化学系,天津市分子光电科学重点实验室,天津300072 [2]西北师范大学化学化工学院,甘肃省生物电化学与环境分析重点实验室,兰州730070

出　　处：《分析化学》2023年第1期53-62,I0001-I0006,共16页Chinese Journal of Analytical Chemistry

基　　金：国家自然科学基金项目(Nos.22174110,22127803,21904095);甘肃省高等学校产业支撑计划项目(No.2021cyzc-01);天津市科技重大专项与工程项目(No.19ZXYXSY00090);中央政府引导地方科技发展专项资金项目(No.2020-2060503-17)资助。

摘　　要：结构特异性核酸酶瓣状核酸内切酶1(Flap enduclease 1,FEN1)在多种癌细胞中过度表达,被认为是癌症临床诊断的潜在生物标志物。FEN1的精确检测对于癌症早期诊断和预后具有重要意义。荧光法具有操作简单、灵敏度高等优势,被广泛应用于FEN1检测,但基于比率荧光信号检测FEN1的策略尚未见报道。本研究设计了一种基于DNA功能化金属有机框架材料(MOFs)的比率荧光探针(FAM-RhB@UiO-66-NH2),用于细胞内FEN1的检测和成像。其中,罗丹明B(RhB)被封装在MOFs中,荧光团FAM修饰的DNA链通过Zr—O—P相互作用与MOFs结合,形成基于荧光共振能量转移(FRET)的纳米探针。在FEN1存在的情况下,DNA链被酶切割,FRET体系被破坏,荧光比率发生变化,从而可实现对FEN1的检测。在最佳实验条件下,检测FEN1的线性范围为0.01~3.0 U,线性方程为y=0.1314x+1.6107,检出限低至0.004 U(3σ)。将此探针用于细胞内FEN1成像,能够区分癌细胞和正常细胞。Structure-specific nuclease(FEN1)is overexpressed in a variety of cancer cells and is considered as a potential biomarker for cancer diagnosis.Accurate detection of FEN1 is of great significance for the early diagnosis and prognosis of cancer.Fluorescence strategies have been widely used in the detection of FEN1 in recent years due to their simple operation and high sensitivity,but the strategy for detecting FEN1 based on ratiometric fluorescence signals has not been reported.Here,a ratiometric fluorescent probe(FAM-RhB@UiO-66-NH2)based on DNA-functionalized metal-organic frameworks(MOFs)was designed for the first time for detection and imaging of intracellular FEN1.Among them,Rhodamine B(RhB)was encapsulated into MOFs,and the fluorophore FAM-modified DNA strand was combined with MOFs through Zr-O-P interaction to form a nanoprobe based on fluorescence resonance energy transfer(FRET).In the presence of FEN1,the DNA strand was cleaved by the enzyme to induce the destruction of the FRET system,resulting in the change of the fluorescence ratio to achieve the detection of FEN1.Under the optimal experimental conditions,the linear range for detection of FEN1 was 0.01-3.0 U,the linear equation was y=0.1314x+1.61071,and the detection limit was as low as 0.004 U(3σ).By using FAM-RhB@UiO-66-NH2as probe,intracellular imaging of FEN1 could distinguish cancer cells from normal cells.

关 键 词：荧光传感 金属有机框架材料 瓣状核酸内切酶1 脱氧核糖核酸 检测 成像 

分 类 号：O657.3[理学—分析化学] R730.43[理学—化学]