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Updated: Aug 31, 2026

Chemical Triphosphorylation of Oligonucleotides
Published on: June 2, 2022
An unnatural base pair for efficient incorporation of nucleotide analogs into RNAs
Ichiro Hirao1, Tsuneo Mitsui, Michiko Kimoto
1Research Center for Advanced Science and Technology, The University of Tokyo, 4-6-1 Komaba, Meguro-ku, Tokyo 153-8904, Japan.
Abstract:
An unnatural base, 2-amino-6-(2-thiazolyl)purine (denoted as v), was developed as an efficient complementary base of another unnatural base, 2-oxo(1H)pyridine (denoted as y). The nucleoside derivatives of v and DNA fragments containing v were chemically synthesized. The efficiency and selectivity of the v-y pairing in replication and transcription were examined and compared to those of a previously developed unnatural base pair between 2-amino-6-(2-thienyl)purine (s) and y. The nucleoside 5'-triphosphate of y (dyTP or yTP) was selectively and efficiently incorporated into DNA or RNA opposite v. The efficiency of the v-y pairing was much higher than that of the s-y pairing. The unnatural v-y pair could be useful for large-scale preparations of RNA molecules containing unnatural bases at desired positions for the expansion of the genetic alphabet.
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