Development of a Structurally Interacting RNA (sxRNA) technology

结构相互作用 RNA (sxRNA) 技术的开发

• 批准号: 10372275
• 负责人: SCOTT A TENENBAUM
• 金额: $ 31.98万
• 依托单位: SUNY POLYTECHNIC INSTITUTE
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-01-01 至 2023-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10372275
• 关键词: Antiviral Agents; Binding; Binding Proteins; Biomanufacturing; Cells; Code; Custom; Development; Ectopic Expression; Engineering; Eukaryotic Initiation Factor-4E; Family; Future; Genes; Genetic Transcription; Genetic Translation; Goals; Grant; Histones; Human Herpesvirus 4; IS Elements; Internal Ribosome Entry Site; Iron; Location; Messenger RNA; MicroRNAs; Molecular; Mutate; Nucleotides; Open Reading Frames; Positioning Attribute; Protein Binding Domain; Proteins; Public Health; RNA; RNA Binding; RNA-Binding Proteins; Resources; Response Elements; Scheme; Selenocysteine; Sensitivity and Specificity; Structure; System; Technology; Testing; Time; Tissues; Translations; Untranslated RNA; Viral; Work; aptamer; base; bioimaging; circular RNA; design; functional restoration; hammerhead ribozyme; interest; piRNA; prevent; protein B; prototype; recruit; sensor; stem; stem cell differentiation; technology research and development; technology validation; tool

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.

项目概要 sxRNA 技术在概念上基于一种 RNA 分子与另一种 RNA 的结合 分子以可预测地“切换”第一个 RNA 的结构确认的方式。作为一个 为了展示 sxRNA 技术的潜力，我们使用 microRNA 的存在/不存在作为 “触发”打开异位传递的 Bait-mRNA 序列的翻译活性，以创建 内源性 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，mRNA 的翻译被激活以产生任何感兴趣的蛋白质。因此 sxRNA 可用于 基于独特表达的存在而异位递送的 mRNA 的瞬时表达 微小RNA。 当前的 sxRNA 技术已基于模块化设计简化为实践，其中包括 可切换的诱饵-RNA 结构（目标 1）、触发-RNA（目标 2）和束缚的开放阅读框编码 寻找感兴趣的基因（目标 3）。这项重点技术研发的长期目标 赠款旨在充分开发 sxRNA 技术，使其能够为公众发挥最广泛的潜力 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展其作为新技术的潜力 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 microRNA 表达作为干细胞分化和制造中靶向细胞/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。

项目成果

PTSelect™: A post-transcriptional technology that enables rapid establishment of stable CHO cell lines and surveillance of clonal variation.

• DOI: 10.1016/j.jbiotec.2020.09.025
• 发表时间: 2021-01-10
• 期刊: Journal of biotechnology
• 影响因子: 4.1
• 作者: Muralidharan-Chari V;Wurz Z;Doyle F;Henry M;Diendorfer A;Tenenbaum SA;Borth N;Eveleth E;Sharfstein ST
• 通讯作者: Sharfstein ST