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

描述（由申请人提供）：1993年发现的小调节RNA分子告诉我们，分子生物学的中心法则比最初认为的要复杂得多。在构建蛋白质的过程中，专门的RNA分子似乎不仅仅是充当中间信使，而是可以通过转录后调控微妙地塑造细胞表型。随后的研究表明，miRNA分子参与了关键的调节网络，例如分化、应激反应和肿瘤发生。被称为微小RNA（miRNAs）的这些非常小（大约17-26个核苷酸）的分子从分子生物学的角度难以操纵。本文讨论的工作将开发和商业化一种独特的分子生物学技术，显着简化microRNA的鉴定和定量。该方法利用了miRNA被RNase III酶加工的事实，RNase III酶在其切割产物上留下特征指纹，可以用适当的分子诱饵回收。报告了先前的I期研究结果，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，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经验的实验室科学家。这项研究工作将通过为研究人员提供下一代miRNA发现收费来影响公共卫生。目前，学术和工业实验室都在努力发现miRNAs及其特定功能。这项研究代表了一个巨大的机会，以提高我们对发育，细胞增殖，分化，细胞周期和疾病（例如，癌症和病毒感染）。根据SBIR开发的产品将使广泛的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.