A DNA structure-mediated fluorescent biosensor for apurinic/apyrimidinic endonuclease 1 activity detection with ultra-high sensitivity and selectivity
A DNA structure-mediated fluorescent biosensor for apurinic/apyrimidinic endonuclease 1 activity detection with ultra-high sensitivity and selectivity
复制标题
一种DNA结构介导的荧光生物传感器，用于检测脱嘌呤/脱嘧啶核酸内切酶1活性，具有超高灵敏度和选择性
DOI：
10.1016/j.snb.2020.129332
复制
发表时间：
2020-12
期刊：
影响因子：
--
通讯作者：
Tongbo Wu
中科院分区：
文献类型：
--
作者：
Yuqiang Hu;Zhen Zhang;Weicong Ye;Wei Zhang;Minghao Hu;Wenqian Yuan;Hongbo Wang;Xianjin Xiao;Tongbo Wu
人脱嘌呤/脱嘧啶核酸内切酶1（APE 1）在DNA修复和基因调控中起着至关重要的作用。人体血液和组织细胞中APE 1浓度的异常变化与多种疾病密切相关。因此，APE 1可用作生物标志物以辅助疾病的临床诊断。同时具有高灵敏度和选择性的APE 1活性的检测是难以实现的。本文提出了一种DNA结构介导的荧光生物传感器来解决上述问题。在APE 1存在时，无嘌呤/无嘧啶位点将被裂解，发夹DNA底物的末端结构将改变。然后，末端脱氧核苷酸转移酶和核酸内切酶IV可提供双重荧光信号扩增。通过巧妙的结构设计，提高了APE 1的反应活性，减少了其他酶的干扰。结果表明，APE 1的检测限低至1.7 × 10− 6 U/mL。该传感器已成功应用于真实的生物样品中APE 1的检测和APE 1抑制剂的筛选。
Human apurinic/apyrimidinic endonuclease 1 (APE1) plays a crucial role in DNA repair and gene regulation. The abnormal variations of the concentration of APE1 in the human blood and tissue cells are highly correlated to various diseases. Thus, APE1 can be used as a biomarker to aid clinical diagnosis of diseases. Detection of APE1 activity with high sensitivity and selectivity simultaneously is difficult to achieve. Here we provide a DNA structure-mediated fluorescent biosensor to solve the above problem. Upon the existence of APE1, the apurinic/apyrimidinic site will be cleaved, and the terminal structure of the hairpin DNA substrate will change. Then the terminal deoxynucleotidyl transferase and endonuclease IV could provide dual fluorescent signal amplification. Through the ingenious structure design of the biosensor, we have improved the reactivity of APE1 and minimized the interference of other enzymes. The results indicate that the detection limit for APE1 is as low as 1.7 × 10−6U/mL. The biosensor has been successfully applied to the detection of APE1 in real biological samples and the screening of APE1 inhibitors.
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