A novel self-feedback biosensor for label-free detection of APE1 by primer exchange reaction and rolling circle amplification/dimeric G-quadruplex

作者全名："Qi, Yinxiao; Du, Yumin; Kang, Qi; Yang, Xiaoyan; Xiang, Hua"

作者地址："[Qi, Yinxiao; Du, Yumin; Xiang, Hua] Chongqing Med Univ, Coll Lab Med, Key Lab Med Diagnost, Minist Educ, Chongqing 400016, Peoples R China; [Kang, Qi] Henan Univ, Huaihe Hosp, Dept Nucl Med, Kaifeng, Peoples R China; [Yang, Xiaoyan] Shandong Univ, Qilu Hosp, Dezhou Hosp, Dezhou, Shandong, Peoples R China"

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

来源：MICROCHEMICAL JOURNAL

ESI学科分类：CHEMISTRY

WOS号：WOS:001067551200001

JCR分区：Q1

影响因子：4.9

年份：2023

卷号：193

期号：　

开始页：　

结束页：　

文献类型：Article

关键词：Human apurinic/apyrimidinic endonuclease 1; (APE1); Primer exchange reaction; G4 dimer; Fluorescent biosensor; Cascade amplification

摘要："Human apurinic/apyrimidinic endonuclease 1(APE1), a critical DNA base excision repair enzyme, plays a fundamental role in nucleotide damage repair and transcriptional regulation, emerging as a vital target for anticancer therapy. Therefore, sensitive and label-free detection of APE1 is necessary. Herein, a primer exchange reaction (PER) cascade rolling circle amplification (RCA)/ dimeric G-quadruplex (G4) self-feedback biosensor was developed to detect APE1. Initially, a gated hairpin (GHP) was meticulously crafted for the specific recognition of APE1, in which GHP is not only the recognition substrate but also acts as a catalytic hairpin (CHP) and a primer for PER. The target recognizes and cleaves the GHP, instigating the PER cycle and engendering extended single-strands DNA, thereby activating RCA reaction. Subsequently, the resulting protracted tandem Grich ssDNA conformationally adapts into a pronounced G4 dimeric structure through affinity binding to K+ and thioxlavin T (ThT), culminating in a stronger fluorescence signal. The design of GHP and G4 dimer greatly improves the signal-to-noise ratio and detection sensitivity, with a wide linear range (0.02-30 U/mL) and a low limit of detection (0.001 U/mL). Moreover, this protocol exhibits robust resistance to interference and holds significant potential in pharmaceutical evaluation, which may provide a reliable analytical method for the detection of APE1 and the subsequent clinical assessment of associated malignancies."

基金机构："Innovative Research Funds for School of Laboratory Medicine, Chongqing Medical University"

基金资助正文："This work was financially supported by Innovative Research Funds for School of Laboratory Medicine, Chongqing Medical University."