Development of high-throughput detection method of tRNA modifications

2015 Fiscal Year Final Research Report

• Project/Area Number: 26670284
• Research Category: Grant-in-Aid for Challenging Exploratory Research
• Allocation Type: Multi-year Fund
• Research Field: Laboratory medicine
• Research Institution: Kumamoto University
• Principal Investigator: Tomizawa Kazuhito 熊本大学, 大学院生命科学研究部(医), 教授 (40274287)
• Project Period (FY): 2014-04-01 – 2016-03-31
• Keywords: 糖尿病 / ミトコンドリア / tRNA / 化学修飾 / 定量PCR

Outline of Final Research Achievements

Modification of tRNAs is a critical factor underlying the development of diabetes, and the measurements of the modification level could be used as a clinical biomarker for the diagnosis of diabetes. However, there is no convenient method to detect tRNA modifications. Current methods, such as mass spectrometry and primer extension method, are not ideal for the easily and reliably detection of such modifications in clinical samples. Here, I present a new method (qPCR-MtR) that overcomes the disadvantages of the current methods and can detect and quantify the modification of tRNAs in multiple samples. The method was sensitive enough to measure 2-methlthio (ms2) modification levels of tRNALys(UUU) in total RNA sample isolated from human peripheral blood specimens and revealed that ms2-modification levels in tRNALys(UUU) were lower in individuals carrying higher-risk SNP of cdkal1.

Free Research Field

生理学