Recognition of unnatural base pairs by a eukaryotic DNA polymerase enables universal sequencing of an expanded genetic alphabet.

ABSTRACT: The expansion of the genetic alphabet through the development of unnatural base pairs (UBPs) has the potential to revolutionize synthetic biology and biotechnology. However, the replication of the UBPs by eukaryotic DNA polymerases is largely unexplored and the sequencing of the UBPs remains challenging. Herein, we explored and demonstrated the activity of human DNA polymerase β (Pol β) for the efficient and specific synthesis and extension of a panel of representative UBPs, including dNaM-dTPT3, dCNMO-dTPT3, and their functionalized derivatives. Based on this, we established a method for the sequencing of DNAs containing different unnatural bases, involving stalled primer extension mediated by Pol β, selective conversion of an unnatural nucleotide into two different natural ones in parall

SUBMITTER: Luo H 

PROVIDER: S-EPMC12781883 | biostudies-literature | 2026 Jan

REPOSITORIES: biostudies-literature