Enzymatic synthesis of site specifically isotope labelled RNA

2016 Fiscal Year Final Research Report

• Project/Area Number: 15K13738
• Research Category: Grant-in-Aid for Challenging Exploratory Research
• Allocation Type: Multi-year Fund
• Research Field: Bio-related chemistry
• Research Institution: Tokyo Institute of Technology
• Principal Investigator: Seio Kohji 東京工業大学, 生命理工学院, 准教授 (20313356)
• Project Period (FY): 2015-04-01 – 2017-03-31
• Keywords: ヌクレオシド三リン酸 / RNAポリメラーゼ / アプタマー / 光ケージドRNA

Outline of Final Research Achievements

We studied the enzymatic incorporation of base- or sugar-modified ribonucleoside triphosphates by T7-RNA polymerase to develop the future technology to synthesize isotope-labeled RNA enzymatically. We tested 6-O-nitrobenzylguanosine and 4-O-nitrobenzyluridine triphosphates as the substrates of T7-RNA polymerase. However, it was revealed that both nucleoside triphosphates acted as inhibitor of the polymerase when concentrations of the nucleoside triphosphates were high. On the contrary, as the sugar modified nucleoside triphosphates, we tested 2'-O-carbamoyluridine triphosphate as the substrate or T7-RNA polyerase and found that this modified nucleoside triphosphate could be incorporated into RNA strand enzymatically. This result suggested that the carmaoyl-modified nucleoside triphosphate can be used for the enzymatic synthesis chemically modified functional RNAs such as RNA aptamers.

Free Research Field

核酸有機化学