Sensitive Detection of uracil-DNA Glycosylase Based on AND-gate Triggers
Sensitive Detection of uracil-DNA Glycosylase Based on AND-gate Triggers
复制标题
基于与门触发的尿嘧啶-DNA 糖基化酶的灵敏检测
DOI：
10.1016/j.snb.2022.132174
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发表时间：
2022
影响因子：
8.4
通讯作者：
Tongbo Wu
中科院分区：
文献类型：
--
作者：
Yujie Zhou;Zhiyuan Zhou;Tianci Xie;Yuqiang Hu;Wei Zhang;Changjiang Li;Zhen Zhang;Xianjin Xiao;Tongbo Wu
As a DNA repair enzyme, uracil-DNA glycosylase (UDG) removes uracil bases from DNA and starts the base excision repair pathway to maintain genomic integrity. It is meaningful to detect UDG activity sensitively because abnormal UDG activity is associated with cancer, immunodeficiency and many other diseases. Fluorescent detection methods have been rapidly developed because of their simplicity and sensitivity. However, in these ways, triggers used to initiate fluorescent signals only come from a single source, either the primer or the enzyme. The accidental leakage of the trigger leads to an unwanted background. To suppress the background signal further, we proposed an improved fluorescent strategy based on AND-gate triggers. In this strategy, a uracil-containing DNA oligo could inhibit the archaeal family B DNA polymerase. After UDG cleavage, the polymerase was activated (trigger-1). Meanwhile, an apurinic/apyrimidinic site was formed, then excised by Endonuclease IV, making the uracil-containing oligo become a primer with 3′-OH end (trigger-2) for further amplification. Then, an exponential amplified fluorescent signal could be generated with nicking endonuclease and well-designed templates. As the two triggers worked as an AND-gate, the leakage of any one of them was insufficient to initiate amplification reactions, resulting in low background. This AND-gate triggers based two-step amplification strategy showed a time-dependent output, with a detection limit of 1.5 × 10 −4 U/mL. In addition, this solution also demonstrated a high selectivity toward UDG. Hence, this proposed strategy can be served as a promising platform for UDG activity detection and associated biomedical studies. • A fluorescent strategy for uracil-DNA glycosylase (UDG) detection is developed. • The strategy uses the polymerase and the primer as the AND-gate triggers. • The AND-gate triggers effectively reduce the background. • This strategy shows satisfying sensitivity and selectivity toward UDG.
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影响因子：
4.7
作者：
T. Vollberg;G. Seal;M. Sirover
通讯作者：
T. Vollberg;G. Seal;M. Sirover
影响因子：
11.4
作者：
Kubota, Y;Nash, RA;Lindahl, T
通讯作者：
Lindahl, T
影响因子：
9.7
作者：
Shuang Zhao;Lianyu Yu;Sha Yang;Xiaoqi Tang;Kai Chang;Ming Chen
通讯作者：
Shuang Zhao;Lianyu Yu;Sha Yang;Xiaoqi Tang;Kai Chang;Ming Chen
影响因子：
6.1
作者：
Jianbo Sun;Changjian Li;Yuqiang Hu;Yawen Ding;Tongbo Wu
通讯作者：
Jianbo Sun;Changjian Li;Yuqiang Hu;Yawen Ding;Tongbo Wu
自引物和自模板循环滚环扩增策略用于灵敏检测尿嘧啶-DNA糖基化酶活性
影响因子：
6.2
作者：
Zhang Pingping;Wang Lei;Zhao Haiyan;Xu Xiaowen;Jiang Wei
通讯作者：
Jiang Wei