SBIR Phase I: New generation of RNA interference (RNAi) research tools to study activity of genes inside living cells

SBIR 第一阶段：新一代 RNA 干扰 (RNAi) 研究工具，用于研究活细胞内基因的活性

• 批准号: 1415657
• 负责人: alexey wolfson
• 金额: $ 15万
• 依托单位: Advanced RNA technologies
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-07-01 至 2015-06-30
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1415657&HistoricalAwards=false
• 关键词: SBIR; Phase; New; generation; RNA

The broader impact/commercial potential of this Small Business Innovation Research (SBIR) project is to provide researchers with more powerful and affordable tools to study activity of genes inside the living cells. RNA interference (RNAi) is a Noble Prize-winning technology that allows specific silencing of gene expression inside living cells. This technology quickly became an invaluable tool in study of gene function, functional identification of new genes, and therapeutic development. Currently, however, available RNAi techniques are limited by non-specific effects and high cost. The goal of this proposal is to develop and make available to a scientific community a novel generation of RNAi reagents, based on the use of chemically modified long double-stranded (ds) RNA molecules. The higher potency and specificity of this technology, along with lower costs, will allow researchers to significantly increase the efficacy of their functional genomics studies. The proposed RNAi technology will further the field of functional genomics and move closer toward creating a functional map of the human genome. It is anticipated that this technology has the potential to replace a significant share of currently available RNAi tools and capture a sizable share of the current RNAi-based research tools market, estimated at $120-200M. This SBIR project aims to develop a new generation of more potent and specific RNAi research tools, based on the introduction of long dsRNAs into mammalian cells. While long dsRNA is used successfully for RNAi screening in C.elegans and flies, it can't be used directly in mammalian cells due to the induction of a strong native immune response. Recent studies have demonstrated that long dsRNA has a potent and specific RNAi effect in mammalian cells, which is usually masked by dsRNA-induced toxicity. Introduction of a variety of nucleotide modifications into mRNA (i.e., long structured RNA molecules) prevents development of this innate immune response. The goal is to validate a number of RNA modifications known to block RNA interactions with Toll-receptors in the context of long dsRNA to circumvent the immune response. This will be accomplished by incorporating a number of modified nucleotides into dsRNA, and then testing them for gene silencing potency and specificity, and toxicity (ability to induce the innate immune response) in mammalian cell culture. RNAi reagents based on long dsRNAs are expected to have higher potency and specificity compared to standard siRNA-based reagents, and provide a simpler and more cost effective way of handling RNAi libraries for functional genomic screening.

这项小型企业创新研究（SBIR）项目的更广泛的影响/商业潜力是为研究人员提供更强大且负担得起的工具，以研究活细胞内基因的活动。 RNA干扰（RNAi）是一项高贵的获奖技术，可以使活细胞内的基因表达进行特定的沉默。这项技术很快成为研究基因功能，新基因的功能鉴定以及治疗开发的宝贵工具。 但是，目前，可用的RNAi技术受非特异性效果和高成本的限制。该提案的目的是基于化学修改的长双链（DS）RNA分子，开发并为科学界开发并提供一种新一代的RNAi试剂。该技术的较高效力和特异性以及较低的成本将使研究人员能够显着提高其功能基因组学研究的功效。拟议的RNAi技术将进一步进一步的功能基因组学领域，并靠近创建人类基因组的功能图。预计该技术有可能取代当前可用的RNAi工具，并在当前基于RNAi的研究工具市场中占有相当大的份额，估计为120-2亿美元。这个SBIR项目旨在根据将长DSRNA引入哺乳动物细胞的基础上，开发出新一代的更有效和特定的RNAi研究工具。虽然长dsRNA成功地用于甲状腺菌和果蝇中的RNAi筛选，但由于诱导强大的天然免疫反应，它不能直接在哺乳动物细胞中使用。最近的研究表明，长dsRNA在哺乳动物细胞中具有有效的特异性RNAi效应，通常被DSRNA诱导的毒性掩盖。将各种核苷酸修饰引入mRNA（即长结构的RNA分子）中，可阻止这种先天免疫反应的发展。目的是验证许多已知的RNA修饰，可在长dsRNA的背景下阻止RNA相互作用与电话受体受体相互作用，以规避免疫反应。 这将通过将许多修饰的核苷酸纳入DSRNA来实现，然后测试它们是否在哺乳动物细胞培养中测试基因沉默的效力和特异性以及毒性（能够诱导先天免疫反应的能力）。 与基于标准的SIRNA的试剂相比，基于长DSRNA的RNAi试剂预计将具有更高的效力和特异性，并提供了一种更简单，更具成本效益的方法来处理RNAi库进行功能性基因组筛选。