Novel DNA/RNA-regulated hammerhead ribozymes for biosensor applications

用于生物传感器应用的新型 DNA/RNA 调节的锤头核酶

• 批准号: 580881-2022
• 负责人: Kharma, NawwafN
• 金额: $ 2.19万
• 依托单位: Concordia University
• 依托单位国家: 加拿大
• 项目类别: Alliance Grants
• 财政年份: 2022
• 资助国家: 加拿大
• 起止时间: 2022-01-01 至 2023-12-31
• 项目状态: 已结题
• 来源: https://www.nserc-crsng.gc.ca/ase-oro/Details-Detailles_eng.asp?id=751861
• 关键词: Novel; DNA; RNA; regulated; hammerhead

This research aims to discover and protect (via patent) a novel method for RNA-based translational regulation, one that could provide an alternative to an existing popular method (toehold switching), which is patented. This, in turn, can be used for the development of cell-free, bio-sensors of pathogenic RNAs. The invention of a new method for translational regulation would present a non-trivial technical discovery. It opens the door to applications such as the monitoring and regulation of gene expression in vivo, in addition to the development by the partner company of kits/tools for pathogenic RNA (e.g., HIV and Hepatitis-C) detection, in human-sourced samples. The proposed method utilizes computationally designed trans-acting hammerhead ribozymes (taHHRzs), which would cleave a reporter gene transcript, unless special 'anti-cleaver' RNAs bind to a designated part of the taHHRz. This renders the taHHRz inactive, protecting the reporter gene transcript from cleavage, and allowing it to be translated to a visually detectable protein (or one that results in a detectable colored residue). In sum, the project's goals (both applied and academic) are well-defined; they require the development and optimization of a new method of translational regulation (outlined in the proposal); a fairly detailed plan of action and budget are presented; and the real-world implications of a satisfactory solution are clear and significant.

本研究旨在发现并保护(通过专利)一种基于RNA的翻译调控的新方法，这种方法可以提供现有流行方法(Toehold Switch)的替代方法，该方法已获得专利。这反过来又可用于开发无细胞的致病RNA生物传感器。一种新的翻译调控方法的发明将带来一项不平凡的技术发现。除了合作伙伴公司开发检测人类来源样本中的致病RNA(如艾滋病毒和丙型肝炎)的试剂盒/工具外，它还为监测和调节体内基因表达等应用打开了大门。该方法利用计算设计的反式锤头状核酶(TaHHRzs)，它将切割报告基因转录本，除非特殊的反切割RNA与taHHRz的指定部分结合。这使得taHHRz不活跃，保护报告基因转录本不被切割，并允许它被翻译成可视觉检测到的蛋白质(或导致可检测到有色残基的蛋白质)。总而言之，该项目的目标(无论是应用的还是学术的)都是明确的；它们需要开发和优化一种新的转化性监管方法(在提案中概述)；提出了相当详细的行动计划和预算；令人满意的解决方案的现实影响是明确和重要的。