THRUST: translesion synthesis-driven hierarchical regulation using a template-activator construct for Cas12a activity.

ABSTRACT: The CRISPR/Cas12a system has demonstrated extraordinary capabilities in biosensing and molecular diagnostics, owing to its precise recognition and efficient trans-cleavage ability. However, current crRNA-based Cas12a regulation is relatively crude, requiring costly modifications and multiple components, increasing system complexity. Here, we develop translesion synthesis-driven hierarchical regulation using a template-activator construct for Cas12a activity (THRUST), a powerful and economical Cas12a regulation strategy. This strategy utilizes a bifunctional template-activator construct (TAC) that simultaneously functions as a transcriptional template for T7 RNA polymerase (T7 RNAP) and an activator for Cas12a. T7 RNAP skips the deoxyuridine (dU) lesion while being blocked by the apu

SUBMITTER: Qin L 

PROVIDER: S-EPMC12168957 | biostudies-literature | 2025 Jul

REPOSITORIES: biostudies-literature