文献类型：期刊文献

英文题名：Cell-Free Biosensing Genetic Circuit Coupled with Ribozyme Cleavage Reaction for Rapid and Sensitive Detection of Small Molecules

作者：Li, Wenxing[1,2];Xu, Yiming[3];Zhang, Yue[3];Li, Pengfei[3];Zhu, Xiaoli[2,4];Feng, Chang[1]

机构：[1]Shanghai Univ, Ctr Mol Recognit & Biosensing, Shanghai Engn Res Ctr Organ Repair, Sch Life Sci, Shanghai 200444, Peoples R China;[2]Tongji Univ, Shanghai Peoples Hosp 10, Dept Clin Lab Med, Shanghai 200072, Peoples R China;[3]East China Univ Sci & Technol, Dept Appl Biol, Shanghai 200237, Peoples R China;[4]Shanghai Univ, Shaoxing Inst, Shaoxing 312071, Peoples R China

年份：2023

卷号：12

期号：6

起止页码：1657

外文期刊名：ACS SYNTHETIC BIOLOGY

收录：;WOS:【SCI-EXPANDED(收录号:WOS:001008953100001)】；

基金：This work was supported by the National Natural Science Foundation of China (grant nos. 32150011 and 22074090), the Clinical research project of Shanghai Tenth People's Hospital (YNCR2A005), and the Special project of Science and Technology Plan of Shaoxing Science and Technology Bureau (no. 2020B33004).

语种：英文

外文关键词：genetic circuit; in vitro transcription; ribozyme; IPTG; inhibitory drugs screening; 3D-printedsensor array

摘要：Syntheticbiological systems have been utilized to developa widerange of genetic circuits and components that enhance the performanceof biosensing systems. Among them, cell-free systems are emergingas important platforms for synthetic biology applications. Geneticcircuits play an essential role in cell-free systems, mainly consistingof sensing modules, regulation modules, and signal output modules.Currently, fluorescent proteins and aptamers are commonly used assignal outputs. However, these signal output modes cannot simultaneouslyachieve faster signal output, more accurate and reliable performance,and signal amplification. Ribozyme is a highly structured and catalyticRNA molecule that can specifically recognize and cut specific substratesequences. Here, by adopting ribozyme as the signal output, we developeda cell-free biosensing genetic circuit coupled with the ribozyme cleavagereaction, enabling rapid and sensitive detection of small molecules.More importantly, we have also successfully constructed a 3D-printedsensor array and thereby achieved high-throughput analysis of an inhibitorydrug. Furthermore, our method will help expand the application rangeof ribozyme in the field of synthetic biology and also optimize thesignal output system of cell-free biosensing, thus promoting the developmentof cell-free synthetic biology in biomedical research, clinical diagnosis,environmental monitoring, and food inspection.