I-Corps: Identification of a stable hybrid carrier for large-scale expression of recombinant miRNAs and siRNAs in E. coli

这个i-Corps项目的更广泛的影响/商业潜力集中在开发一种高产量、高成本效益的天然RNA干扰(RNAi)试剂生产平台。这个项目解决了人们对开发用于治疗学和转化性研究的天然RNA试剂的日益增长的兴趣。由于基于RNA的药物具有接近其他方法无法获得的靶点(如小分子和蛋白质)的能力，该项目将为使用天然生产的药物治疗不治之症开辟新的方向。随着发酵技术的使用，这项研究可能会为学术研究机构和制药公司带来具有成本效益的天然RNA药物的生产。如果成功，该项目将建立一个新型的RNAi生物工程平台。这个I-Corps项目是基于鉴定出一种稳定的、完全人性化的杂交载体，用于在一种常见的大肠杆菌菌株中大规模表达重组微型RNA(MiRNA)和小干扰RNA(SiRNA)。目前的RNA干扰(RNAi)研究和开发往往局限于使用化学工程或合成的RNAi试剂，这些试剂需要各种广泛的人工修饰才能稳定。这些人工修饰可以激发免疫原性。该项目的总体目的是开发一个独特的平台，以经济和可持续的方式生产在活细胞中生产和折叠的天然RNAi制剂。早期工作表明，生物工程miRNA/siRNA试剂携带的天然转录后修饰核苷很少，在人类细胞和小鼠模型中表现出良好的稳定性。该奖项反映了NSF的法定使命，并通过使用基金会的智力优势和更广泛的影响审查标准进行评估，被认为值得支持。

The broader impact/commercial potential of this I-Corps project focuses on the development of a high-yield, cost-effective production platform for natural RNA interference (RNAi) agents. This project addresses the increasing interest in developing natural RNA-based agents for therapeutics and translational research. Since RNA-based drugs have the capability to approach targets (such as small molecules and proteins) that are unavailable by other methods, this project will open new directions for treating incurable diseases with naturally produced drugs. With the use of fermentation technology, this research may result in the cost-effective production of natural RNA-based drugs for both academic research institutes and pharmaceutical companies. If successful, this project will establish a novel RNAi bioengineering platform.This I-Corps project is based on the identification of a stable, fully humanized hybrid carrier for large-scale expression of recombinant micro RNA (miRNA) and small interfering RNA (siRNA) in a common strain of E. coli. Current RNA interference (RNAi) research and development is often limited to the use of chemically-engineered or synthesized RNAi agents that require various and extensive artificial modifications for stability. These artificial modifications can provoke immunogenicity. The overall purpose of this project is the development of a unique platform for the economical and sustainable production of natural RNAi agents that are produced and folded in living cells. Early work has demonstrated that bioengineered miRNA/siRNA agents carry few natural post transcriptionally-modified nucleosides and exhibit favorable stability within human cells and mouse models.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.