Site-specific fluorescent labeling of RNA by a base-pair expanded transcription system.
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.
Kawai
Rie
R
Protein Research Group, RIKEN Genomic Sciences Center, 1-7-22 Suehiro-cho, Tsurumi-ku, Yokohama, Kanagawa 230-0045, Japan.
Kimoto
Michiko
M
Mitsui
Tsuneo
T
Yokoyama
Shigeyuki
S
Hirao
Ichiro
I
D020029:Base Pairing; D005456:Fluorescent Dyes / Q000737:chemistry*; D009711:Nucleotides / Q000138:chemical synthesis / Q000737:chemistry; D012313:RNA / Q000737:chemistry*; D014158:Transcription, Genetic
D005456:Fluorescent Dyes; D009711:Nucleotides; D012313:RNA