Site‐Specific Functional Labeling of Nucleic Acids by In Vitro Replication and Transcription using Unnatural Base Pair Systems

Abstract

AbstractExpansion of the genetic alphabet by an artificial extra base pair (unnatural base pair) system allows for the site‐specific functionalization of nucleic acids by polymerase reactions. By attaching functional groups of interest to an unnatural base, a functional unnatural base substrate can be incorporated as a fifth base into DNA or RNA at desired positions, opposite its complementary sixth unnatural base partner in templates. Over the past twenty years, several unnatural base pairs that function in replication and/or transcription in vitro have been developed, and their utility in PCR amplification and T7 transcription has been demonstrated. Here we introduce the development of unnatural base pairs and their application to replication and transcription, with particular focus on our unnatural base pair systems.