dTPT3TP

dTPT3TP is a non-natural nucleoside triphosphate substrate of SFM4‑6 polymerase, which efficiently incorporates at the site corresponding to template dNaM at 50 °C to form a non-natural base pair, supporting primer extension and full-length synthesis of RNA and 2′‑modified RNA oligonucleotides; furthermore, it enables a controllable pause‑restart mode of SFM4‑6 for synthesizing 5′‑end-labeled RNA derivatives^[1].
dTPT3TP and dNaMTP are added together to the culture medium, supporting the stable propagation of plasmids containing the dNaM-dTPT3 unnatural base pair in engineered Escherichia coli strains and enabling the expression of IL-2 protein variants bearing AzK substitutions.^[2].
dTPT3TP can be utilized by SFM4-3 polymerase to synthesize single-stranded DNA bearing dTPT3; dTPT3TP can be recognized by human Pol β and pairs with dNaM and dCNMO on the template, and supports Pol β in completing full-length primer extension, gap filling, and strand displacement extension of dNaM-dTPT3 and dCNMO-dTPT3^[3].
dTPT3TP is incorporated by Pol β, with the highest efficiency opposite dCNMO, with a K_m of 21.91 µM and a k_cat/K_m of 9.58 × 10³ M^-1·min^-1, followed by dNaM, and then dTPT3^[3].