New methods for discovery and study of microRNAs and other small noncoding RNAs

描述(申请人提供)：1993年发现的小调节RNA分子告诉我们，分子生物学的中心教条比最初认为的要复杂得多。似乎特殊的RNA分子可以通过转录后调节巧妙地塑造细胞表型，而不是简单地充当构建蛋白质的中间信使。后来的研究表明，miRNA分子参与了关键的调控网络，如分化、应激反应和肿瘤发生。这些非常小的(大约17-26个核苷酸)分子被称为microRNAs(MiRNAs)，从分子生物学的角度来看很难操纵。本文讨论的工作将开发一种独特的分子生物学技术并将其商业化，该技术大大简化了microRNAs的鉴定和定量。这种方法利用了这样一个事实，即miRNAs是由RNaseIII酶处理的，这在其切割产物上留下了一个特征指纹，可以用适当的分子诱饵进行回收。报告了先前第一阶段研究的结果，其中获得了原理证明和初步结果。第二阶段的提案概述了实验方法，该方法将完善技术，扩大产品组合，并设计产品，将技术直接带给缺乏任何先前miRNA经验的板凳科学家。这项研究工作将通过向研究人员提供下一代miRNA发现通行费来影响公共健康。目前，学术和工业实验室正在进行大量的努力来发现miRNAs及其特定功能。这项研究是一个巨大的机会，可以增进我们对发育、细胞增殖、分化、细胞周期和疾病(如癌症和病毒感染)的了解。在该SBIR下开发的产品将使广大生命科学研究人员能够发现miRNA。

DESCRIPTION (provided by applicant): The 1993 discovery of small regulatory RNA molecules has taught us that the central dogma of molecular biology is much more complicated that originally thought. Rather than simply acting as a middle messenger in the quest to build proteins, it seemed that specialized RNA molecules could subtly mold a cellular phenotype through post- transcriptional regulation. Subsequent research has revealed that miRNA molecules are involved in critical regulatory networks such as differentiation, stress response, and oncogenesis. Known as microRNAs (miRNAs), these very small (approximately 17-26 nucleotides) molecules are difficult to manipulate from a molecular biology perspective. The work discussed herein will develop and commercialize a unique molecular biology technique that significantly simplifies the identification and quantitation of microRNAs. The method takes advantage of the fact that miRNAs are processed by RNaseIII enzymes, which leave a characteristic fingerprint on their cleavage products that can be recovered with the appropriate molecular }bait}. Results of a previous Phase I study are reported, in which proof of principle and initial results were obtained. The Phase II proposal outlines the experimental methodology that will refine the techniques, expand the portfolio, and design products that will bring the technology directly to bench scientists that lack any prior miRNA experience. This research effort will impact public health by providing researches with a next generation miRNA discovery tolls. Currently, there is a substantial effort in academic and industry laboratories to discovery miRNAs, as well as their specific functions. This research represents a tremendous opportunity to enhance our understanding of development, cell proliferation, differentiation, cell cycle, and disease (e.g., cancer and viral infections). The products developed under this SBIR will enable a broad class of life science researchers in miRNA discovery.