A novel biosensor based on tetrahedral DNA nanostructure and terminal deoxynucleotidyl transferase-assisted amplification strategy for fluorescence analysis of uracil-DNA glycosylase activity

作者全名："Zhao, Zixin; Xie, Zuowei; Chen, Siyi; Chen, Min; Wang, Xingyu; Yi, Gang"

作者地址："[Zhao, Zixin; Xie, Zuowei; Chen, Siyi; Chen, Min; Wang, Xingyu; Yi, Gang] Chongqing Med Univ, Coll Lab Med, Key Lab Clin Lab Diagnost, Minist Educ, Chongqing 400016, Peoples R China"

通信作者："Yi, G (通讯作者)，Chongqing Med Univ, Coll Lab Med, Key Lab Clin Lab Diagnost, Minist Educ, Chongqing 400016, Peoples R China."

来源：ANALYTICA CHIMICA ACTA

ESI学科分类：CHEMISTRY

WOS号：WOS:001019303000001

JCR分区：Q1

影响因子：5.7

年份：2023

卷号：1271

期号：　

开始页：　

结束页：　

文献类型：Article

关键词：Uracil DNA glycosylase; Tetrahedral DNA nanomaterials; Copper nanoparticles; Terminal deoxynucleotidyl transferase; Collapse reaction

摘要："Tetrahedral DNA nanostructure (TDN), as a classical bionanomaterial, which not only has excellent structural stability and rigidity, but also possesses high programmability due to strict base-pairs complementation, is widely used in various biosensing and bioanalysis fields. In this study, we first constructed a novel biosensor based on Uracil DNA glycosylase (UDG) -triggered collapse of TDN and terminal deoxynucleotidyl transferase (TDT)-induced insertion of copper nanoparticles (CuNPs) for fluorescence and visual analysis of UDG activity. In the presence of the target enzyme UDG, the uracil base modified on the TDN were specifically identified and removed to produce an abasic site (AP site). Endonuclease IV (Endo.IV) could cleave the AP site, making the TDN collapse and generating 3 & PRIME;-hydroxy (3 & PRIME;-OH), which were then elongated under the assistance of TDT to produce poly (T) sequences. Finally, Copper (II) sulfate (Cu2+) and L-Ascorbic acid (AA) were added to form CuNPs using poly (T) sequences as templates (T-CuNPs), resulting in a strong fluorescence signal. This method exhibited good selectivity and high sensitivity with a detection limit of 8.6 x 10-5 U/mL. Moreover, the strategy has been successfully applied to the screening of UDG inhibitors and the detection of UDG activity in complex cell lysates, which means that it has promising applications in clinical diagnosis and biomedical research."

基金机构：Key Project of Science and Technology Research Program of Chongqing Education Commission [KJZD-K202000404]; Special Fund Project Key Laboratory of Clinical Laboratory Diagnostics (Ministry of Education)

基金资助正文：This work was supported by the Key Project of Science and Technology Research Program of Chongqing Education Commission (Grant No. KJZD-K202000404) and the Special Fund Project Key Laboratory of Clinical Laboratory Diagnostics (Ministry of Education) .