Novel DNA/RNA-regulated hammerhead ribozymes for biosensor applications

本研究旨在发现和保护（通过专利）一种基于rna的翻译调控的新方法，这种方法可以为现有的流行方法（支点转换）提供一种替代方法，该方法已获得专利。反过来，这可以用于开发无细胞的、致病rna的生物传感器。翻译调节新方法的发明将是一项重要的技术发现。它为体内基因表达的监测和调控等应用打开了大门，此外，合作伙伴公司还开发了用于在人类来源样本中检测致病性RNA（例如艾滋病毒和丙型肝炎）的试剂盒/工具。提出的方法利用计算设计的反式作用锤头核酶（taHHRzs），它可以切割报告基因转录物，除非特殊的“抗切割”rna结合到taHHRz的指定部分。这使得taHHRz失去活性，保护报告基因转录物不被切割，并允许它被翻译成视觉上可检测的蛋白质（或导致可检测的彩色残基的蛋白质）。总之，项目的目标（应用的和学术的）是明确的；它们需要开发和优化一种新的翻译监管方法（在提案中概述）；提出了相当详细的行动计划和预算；一个令人满意的解决方案对现实世界的影响是明确而重要的。

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.