Polymerase-Mediated Incorporation of Unnatural Alkynylated Purine–Pyridazine Base Pairs
Abstract
Unnatural base pairs (UBPs) provide a promising strategy for expanding the genetic alphabet; however, their practical application requires efficient recognition by DNA polymerases. Here, we report the polymerase-mediated incorporation of alkynylated purine-pyridazine (Pu-Pz) base pairs bearing pseudo-nucleobase recognition units positioned in the major groove. Single-nucleotide insertion assays revealed that complementary Pu-Pz pairs are selectively incorporated into DNA by KOD DNA polymerase with efficiencies comparable to those of natural base pairs. Furthermore, efficient primer extension was maintained following UBP incorporation, indicating that bulky major-groove modifications do not significantly impede polymerase processivity. These results demonstrate the polymerase compatibility of major-groove-functionalized UBPs and highlight the potential of Pu-Pz pairs as building blocks for expanded genetic systems and site-specific DNA functionalization.
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BibTeX
@article{fan2026polymerasemediated,
title = {{Polymerase-Mediated Incorporation of Unnatural Alkynylated Purine–Pyridazine Base Pairs}},
author = {Wenjue Fan and Hidenori Okamura and Zhuoxin Dong and Giang Hoang Trinh and Yosuke Taniguchi and Fumi Nagatsugi},
journal = {Chemical and Pharmaceutical Bulletin},
year = {2026},
month = jul,
doi = {10.1248/cpb.c26-00263},
url = {https://www.jstage.jst.go.jp/article/cpb/74/7/74_c26-00263/_article},
note = {CC BY-NC},
}
RIS
TY - JOUR
TI - Polymerase-Mediated Incorporation of Unnatural Alkynylated Purine–Pyridazine Base Pairs
AU - Wenjue Fan
AU - Hidenori Okamura
AU - Zhuoxin Dong
AU - Giang Hoang Trinh
AU - Yosuke Taniguchi
AU - Fumi Nagatsugi
PY - 2026
DA - 2026/07/22
JO - Chemical and Pharmaceutical Bulletin
DO - 10.1248/cpb.c26-00263
UR - https://www.jstage.jst.go.jp/article/cpb/74/7/74_c26-00263/_article
AB - Unnatural base pairs (UBPs) provide a promising strategy for expanding the genetic alphabet; however, their practical application requires efficient recognition by DNA polymerases. Here, we report the polymerase-mediated incorporation of alkynylated purine-pyridazine (Pu-Pz) base pairs bearing pseudo-nucleobase recognition units positioned in the major groove. Single-nucleotide insertion assays revealed that complementary Pu-Pz pairs are selectively incorporated into DNA by KOD DNA polymerase with efficiencies comparable to those of natural base pairs. Furthermore, efficient primer extension was maintained following UBP incorporation, indicating that bulky major-groove modifications do not significantly impede polymerase processivity. These results demonstrate the polymerase compatibility of major-groove-functionalized UBPs and highlight the potential of Pu-Pz pairs as building blocks for expanded genetic systems and site-specific DNA functionalization.
SN - 1347-5223
SN - 0009-2363
ER -