新型基因表达调控元件——人工核糖开关的构建及筛选

Engineering and screening of artificial riboswitch as a novel gene control element

摘要:

核糖开关作为一种新发现的RNA元件，可以高效、准确、快速地执行基因调控任务，且免疫原性低，有可能在将来以顺式模块的方式应用于未来的基因治疗。近年来已经成功构建了多种人造核糖开关，构建方法主要是利用人工适体元件与基因表达调控元件组装，或者是在天然核糖开关基础上进行改造。文中全面综述了涉及人工核糖开关设计及筛选的技术，讨论了可以用于哺乳细胞、响应非天然配体信号、调控特征为热力学和动力学控制的核糖开关的设计新策略，并对核糖开关的筛选构建策略及其在基因治疗及新型药物开发领域的应用前景进行了展望。尽管目前将核糖开关设计成为功能强大的新型基因调控系统还面临很大的困难，但通过构效关系的研究、计算机辅助设计、体外筛选及细胞内筛选技术、高通量优化筛选等技术的综合应用，核糖开关一定可以成为有力的基因调控工具，如能成功应用则可大大促进基因治疗临床化的进程。

Abstract:

Various artificial riboswitches have been constructed by utilization of designed aptamers or by modification of natural riboswitch systems, because they can regulate gene expression in a highly efficient, precise and fast way, and promise to supply simple cis-acting, modular, and non-immunogenic system for use in future gene therapy applications. In this review, we present an overview of currently available technologies to design and select engineered riboswitches, and discuss some possible technologies that would allow them highly responsive to non-natural ligands, and dynamic control of gene expression in mammalian cells. Though how to bring custom-designed riboswitches as a novel and versatile tool box to gene control system is still a great challenge, the combination of structure-activity relationship information, computer based molecular design, in vitro selection, and high-through screening will serve as powerful tools for further development of riboswitch based gene regulatory systems.