Site-specific biotinylation of RNA molecules by transcription using unnatural base pairs.
Direct site-specific biotinylation of RNA molecules was achieved by specific transcription mediated by unnatural base pairs. Unnatural base pairs between 2-amino-6-(2-thienyl)purine (denoted by s) and 2-oxo(1H)pyridine (denoted by y), or 2-amino-6-(2-thiazolyl)purine (denoted as v) and y specifically function in T7 transcription. Using these unnatural base pairs, the substrate of biotinylated-y (Bio-yTP) was selectively incorporated into RNA, opposite s or v in the DNA templates, by T7 RNA polymerase. This method was applied to the immobilization of an RNA aptamer on sensor chips, and the aptamer accurately recognized its target protein. This direct site-specific biotinylation will provide a tool for RNA-based biotechnologies.
Moriyama
Kei
K
Research Center for Advanced Science and Technology, The University of Tokyo 4-6-1 Komaba, Meguro-ku, Tokyo 153-8904, Japan.
Kimoto
Michiko
M
Mitsui
Tsuneo
T
Yokoyama
Shigeyuki
S
Hirao
Ichiro
I
D020029:Base Pairing; D001710:Biotin / Q000031:analogs & derivatives* / Q000138:chemical synthesis / Q000737:chemistry / Q000378:metabolism; D019921:Biotinylation; D012321:DNA-Directed RNA Polymerases / Q000378:metabolism; D009705:Nucleosides / Q000138:chemical synthesis / Q000737:chemistry* / Q000378:metabolism; D009843:Oligoribonucleotides / Q000737:chemistry*; D011685:Purine Nucleotides / Q000737:chemistry; D011687:Purines / Q000737:chemistry; D011728:Pyridones / Q000737:chemistry; D012313:RNA / Q000096:biosynthesis / Q000737:chemistry*; D020349:Surface Plasmon Resonance; D013698:Templates, Genetic; D014158:Transcription, Genetic; D014764:Viral Proteins / Q000378:metabolism
C435686:2-amino-6-(2-thienyl)purine; C503404:4-(3-(biotinamido)propynyl)-2-oxo(1H)pyridin-3-ylribofuranosyl triphosphate; D009705:Nucleosides; D009843:Oligoribonucleotides; D011685:Purine Nucleotides; D011687:Purines; D011728:Pyridones; D014764:Viral Proteins; D012313:RNA; D001710:Biotin; C062210:bacteriophage T7 RNA polymerase; D012321:DNA-Directed RNA Polymerases