Development of a Structurally Interacting RNA (sxRNA) technology

项目总结 SxRNA技术在概念上是基于一个RNA分子与另一个RNA分子的结合 分子以一种可预见的方式“切换”第一个RNA的结构确认。作为一名 展示了sxRNA技术的潜力，我们使用了microRNA的存在/不存在作为 “Trigger”打开异位传递的诱饵-mRNA序列的翻译活性，以创建 内源RNA结合蛋白(RBP)的结合基序。一旦RBP被招募到现在的职能部门 基序，信使核糖核酸的翻译被激活以产生任何感兴趣的蛋白质。因此，sxRNA可用于 基于存在差异表达的异位递送mRNA的瞬时表达 MicroRNA。 目前的sxRNA技术已经简化为基于模块化设计的实践，该模块化设计包括 可切换的诱饵-RNA结构(Aim 1)、触发-RNA(Aim 2)和系留开放阅读框编码 寻找感兴趣的基因(目标3)。这项重点技术研发的长期目标 格兰特是为了充分开发sxRNA技术，使其能够在公众中发挥最大的潜力 健康。相应地，这些目标旨在促进全面发展、优化和 对现有sxRNA技术的每个模块进行改进，并将其作为新的潜力进行扩展 基于RNA的分子资源，具有多种应用，包括作为实时的单细胞传感器 MicroRNA表达作为靶向细胞/组织选择的工具，在干细胞分化和制造中， 用于生物制造，作为一种抗病毒/细菌技术和一种新的基于RNA的细胞和组织工具 生物成像。

PROJECT SUMMARY The sxRNA Technology is conceptually based on the binding of one RNA molecule to a second RNA molecule in a manner that predictably “switches” the structural confirmation of the first RNA. As a demonstration of the potential of sxRNA technology, we have used the presence/absence of a microRNA as a “trigger” to turn ON the translational activity of an ectopically delivered Bait-mRNA sequence to create the binding-motif for an endogenous RNA-Binding Protein (RBP). Once the RBP is recruited to the now functional motif, translation of the mRNA is activated to produce any protein of interest. Thus sxRNA can be used for the transient expression of an ectopically delivered mRNA based on the presence of distinctively expressed microRNAs. The current sxRNA technology has been reduced to practice based around a modular design comprised of the switchable bait-RNA structure (Aim 1), the trigger-RNA (Aim 2) and the tethered open reading frame coding for a gene of interest (Aim 3). The long-term objective of this Focused Technology Research and Development grant is to fully develop the sxRNA technology so that it can be utilized to its broadest potential for Public Health. Accordingly, the Aims are designed to facilitate the comprehensive development, optimization and improvement of each of the modules of the existing sxRNA technology and expand the potential as a new RNA-based molecular resource with multiple applications including as a real-time, single-cell sensor of microRNA expression, as a tool for targeted cell/tissue selection, in stem-cell differentiation and manufacture, for biomanufacturing, as an anti-viral/bacterial technology and as a new RNA-based tool for cell and tissue bioimaging.