Amplification-free miRNA detection with CRISPR/Cas12a system based on fragment complementary activation strategy.

ABSTRACT: CRISPR/Cas12a systems have been repurposed as powerful tools for developing next-generation molecular diagnostics due to their trans-cleavage ability. However, it was long considered that the CRISPR/Cas12a system could only recognize DNA targets. Herein, we systematically investigated the intrinsic trans-cleavage activity of the CRISPR/Cas12a system (LbCas12a) and found that it could be activated through fragmented ssDNA activators. Remarkably, we discovered that the single-stranded DNA (ssDNA) activators in the complementary crRNA-distal domain could be replaced by target miRNA sequences without the need for pre-amplification or specialized recognition mechanisms. Based on these findings, we proposed the "Fragment Complementary Activation Strategy" (FCAS) and designed revers

SUBMITTER: Zhao S 

PROVIDER: S-EPMC11495492 | biostudies-literature | 2024 Oct

REPOSITORIES: biostudies-literature