Development of a Structurally Interacting RNA (sxRNA) technology

项目摘要 sxRNA技术在概念上基于一个RNA分子与第二个RNA的结合 在一些实施方案中，第一RNA以可预测地“切换”第一RNA的结构确认的方式与第二RNA结合。作为 为了证明sxRNA技术的潜力，我们使用microRNA的存在/不存在作为 “触发”以开启异位递送的诱饵-mRNA序列的翻译活性，从而产生特异性的诱饵-mRNA序列。 内源性RNA结合蛋白（RBP）的结合基序。一旦RBP被招募到现在的功能 基序，mRNA的翻译被激活以产生任何感兴趣的蛋白质。因此，sxRNA可以用于 基于特异性表达的存在，异位递送的mRNA的瞬时表达 microRNAs。 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： 可切换诱饵RNA结构（Aim 1）、触发RNA（Aim 2）和束缚的开放阅读框架编码 目的基因（Aim 3）。本重点技术研发的长期目标 赠款是为了充分发展sxRNA技术，使其能够利用其最广泛的潜力，为公众 健康因此，目标旨在促进全面发展、优化和 改进现有sxRNA技术的每个模块，并扩大其作为新的 基于RNA的分子资源，具有多种应用，包括作为实时单细胞传感器， 在干细胞分化和制造中，microRNA表达作为靶向细胞/组织选择的工具， 用于生物制造，作为抗病毒/细菌技术，以及作为细胞和组织的新的基于RNA的工具， 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.