核糖核酸调节子的构建与即时检测中的应用  

Construction of Riboregulator in the Application of Point-of-care Testing

作　　者：梁书瑞 李娇娇 齐浩 LIANG Shu-rui;LI Jiao-jiao;QI Hao(Synthetic Biology Research Platform,Tianjin Collaborative Innovation Center of Chemical Science and Engineering,Key Laboratory of Systems Bioengineering of Ministry of Education,School of Chemical Engineering and Technology,Tianjin University,Tianjin 300072,China)

机构地区：[1]天津大学化工学院系统生物工程教育部重点实验室,天津化学化工协同创新中心合成生物学平台,天津300072

出　　处：《中国生物工程杂志》2022年第9期67-82,共16页China Biotechnology

基　　金：国家自然科学基金(21778039、21621004);国家重点研发计划(2019YFA0904103)资助项目。

摘　　要：合成生物学专注于可重复利用模块和元件的工程化设计,并在生物系统中表现出良好的行为和功能。在无细胞蛋白表达系统中,核糖核酸调节子作为即时检测中的重要传感元件,通过靶标分子的诱导使其自身结构发生变化,进而调控下游基因的表达。系统介绍不同类型的核糖核酸调节子及其作用原理,包括一代核糖核酸调节子、toehold开关、功能拓展的核糖核酸调节子和核糖开关。详细阐明构建核糖核酸调节子的设计-测试-分析过程:计算机辅助设计、基因表达测试和结构功能化分析。汇总基于核糖核酸调节子的体外即时检测应用,重点总结toehold开关介导的病原菌核酸检测和核糖开关参与的小分子检测。讨论当前无细胞即时检测的特点、挑战和发展趋势,为开发新型核糖核酸调节子和即时检测工具提供思路和参考。Ongoing efforts in synthetic biology are focused on the design of reusable and modular fragments that demonstrate well-characterized behaviors and functionalities in biological systems. In the cell-free protein expression system, riboregulators are developed as an important sensing element in point-of-care testing, which can change their structure through the induction of target molecules, thereby regulating the expression of downstream genes. Different types of riboregulators and their action mechanisms are systematically introduced, including traditional riboregulators, toehold switch, toehold repressor, three-way junction repressor, small transcription activating RNA, and riboswitch. The main processes of constructing riboregulators are described in detail: designing with computers, testing gene expression, and analyzing the functional structure. Finally, the applications of in vitro point-of-care testing(POCT) based on riboregulators are summarized, mainly including pathogen nucleic acid detection based on toehold switch and small molecule detection mediated by riboswitch. The characteristics, challenges, and development tendencies of POCT in the cell-free system are discussed and summarized. In conclusion, the construction and application of riboregulators have opened up a new direction in the biosensing field. This article is expected to provide some helpful insight into the development of new riboregulators and POCT tools.

关 键 词：核糖核酸调节子 Toehold开关 核糖开关 无细胞蛋白表达 即时检测 

分 类 号：Q78[生物学—分子生物学]