Sequencing Transfer RNAs by Mass Spectrometry

分析和表面化学计划，与分子和细胞生物科学基因和基因组系统集群共同资助，支持辛辛那提大学的帕特里克林巴赫教授的工作，目标是基于质谱的方法来更高通量地测序转移核糖核酸（tRNA）。许多类型的RNA参与所有活细胞中的关键细胞功能。在所有已知的活细胞内的生物有机分子中，RNA分子是唯一存储遗传信息并充当催化剂的分子。在这项研究的过程中，Limbach博士和他的团队将建立新的方法来破碎tRNA的混合物;将开发一种tRNA富集策略，允许稀有tRNA表达和测序;将使用正常和有限的RNase消化的组合来增加tRNA测序读长;并将增强签名消化产物的RNA数据库。该项目将为研究人员提供额外的工具，可以检查转录和翻译过程，补充现有的基因组和蛋白质组学策略。这些发展将为RNA研究人员提供有利的进展，并可转化为核酸分析的相关领域（例如，除了上述技术影响外，这项工作还通过为学生提供生物质谱关键领域的教育和培训，产生了更广泛的影响。林巴赫博士致力于扩大在科学领域代表性不足的少数民族的数量，特别是质谱。

The Analytical and Surface Chemistry Program, with co-funding from the Molecular and Cellular Biosciences Genes and Genome Systems Cluster, supports Prof. Patrick Limbach at the University of Cincinnati for work targeting a mass spectrometry-based approach to higher throughput sequencing of transfer ribonucleic acids (tRNAs). Many types of RNAs are involved in key cellular functions in all living cells. Among all known bio-organic molecules within living cells, RNA molecules are the only ones that store genetic information and act as catalysts. In the course of this research, Dr. Limbach and his group will establish new methods to fractionate mixtures of tRNAs; will develop a strategy for tRNA enrichment allowing rare tRNAs to be expressed and sequenced; will use a combination of normal and limited RNase digestion for increasing tRNA sequencing read length; and will enhance the RNA database of signature digestion products. The project will provide researchers with additional tools that can examine transcription and translation processes, complementing existing genomic and proteomic strategies. These developments will provide enabling advances for RNA researchers and can be translated to related areas of nucleic acid analysis (e.g., xenobiotic modifications).In addition to the technical impacts outlined above, the work offers broader impacts by providing students with education and training in key areas of biological mass spectrometry. Dr. Limbach strives to expand the number of underrepresented minorities engaged in science in general, and mass spectrometry in particular.