High Fidelity Enzyme-Free Primer Extension with an Ethynylpyridone Thymidine Analog
J. Han et.al. 2021 Chemistry https://doi.org/10.1002/chem.202102996
24.09.2021
Jianyang Han, Eric Kervio, and Clemens Richert
Chemistry https://doi.org/10.1002/chem.202102996
Abstract
High fidelity base pairing is important for the transmission of genetic information. Weak base pairs can lower fidelity, complicating sequencing, amplification and replication of DNA. Thymidine 5'-monophosphate (TMP) is the most weakly pairing nucleotide among the canonical deoxynucleotides, causing high errors rates in enzyme-free primer extension. Here we report the synthesis of an ethynylpyridone C -nucleoside analog of 3'-amino-2',3'-dideoxythymidine monophosphate and its incorporation in a growing strand by enzyme-free primer extension. The ethynylpyridone C -nucleotide accelerates extension more than five-fold, reduces misincorporation and readily displaces TMP in competition experiments. The results bode well for the use of the C -nucleoside as replacements for thymidine in practical applications.