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Nucleic Acids Symposium Series 2004 48(1):35-36; doi:10.1093/nass/48.1.35
© 2004 by Oxford University Press
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Site-specific fluorescent labeling of RNA by a base-pair expanded transcription system

Rie Kawai1, Michiko Kimoto1, Tsuneo Mitsui1, Shigeyuki Yokoyama1,3,4 and Ichiro Hirao1,2

1 Protein Research Group, RIKEN Genomic Sciences Center, 1-7-22 Suehiro-cho, Tsurumi-ku, Yokohama, Kanagawa 230-0045, Japan, 2 Research Center for Advanced Science and Technology, The University of Tokyo, 4-6-1 Komaba, Meguro-ku, Tokyo 153-8904, Japan, 3 Department of Biophysics and Biochemistry, Graduate School of Science, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-0033, Japan, 4 RIKEN Harima Institute at Spring-8, 1-1-1 Kohto, Mikazuki-cho, Sayo, Hyogo 679-5148, Japan

Ribonucleoside 5'-triphosphates of fluorescently-labeled 2-oxo(1H)pyridines, in which carboxyfluorescein (FAM) or carboxytetramethylrhodamine (TAMRA) was attached at position 5, were chemically synthesized. These substrates were site-specifically incorporated into RNA by T7 transcription mediated by unnatural base pairs between 2-amino-6-(2-thienyl)purine and 2-oxo(1H)pyridine or 2-amino-6-(2-thiazolyl)purine and 2-oxo(1H)pyridine. This site-specific fluorescent labeling of RNA fragments will be a powerful tool for the fields of nucleic acid chemistry, biology, and technology.


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