Universal Regulation of DNA Polymerase via Photocaged Primer Enables Light-Start Isothermal Amplification on Demand.

ABSTRACT: The development of DNA polymerases with controllable functions, such as hot-start or light-start variants, allows on-demand activation of enzymatic activity and therefore improves reaction specificity. However, the fine control of DNA polymerase activity faces challenges to accuracy and flexibility. Here, a simple and general photoactivatable approach for the temporal regulation of DNA polymerase activity, and thus, light-start isothermal amplification on demand is engineered. This approach leverages photocaged primer with light-sensitive 6-nitropiperonyloxymethyl to temporarily inhibit the conformational change of DNA polymerase until reactivation by 365 nm UV. The mechanism through biochemical assays and molecular dynamic simulations is elucidated, and validate the temporal precision usi

SUBMITTER: Qing M 

PROVIDER: S-EPMC12806212 | biostudies-literature | 2026 Jan

REPOSITORIES: biostudies-literature