A dual-recognition precise nucleic acid probe based on tetrahedron and Cas12a reveals gene-environment interactions

Abstract

Precise detection of gene expression is crucial for understanding the interactions between organisms and their environment. In this study, we developed a highly sensitive dual-recognition gene detection method based on the CRISPR-Cas12a system and DNA tetrahedron and validated its application in biological samples. By optimizing the Cas12a signal readout system, the method demonstrated excellent sensitivity and stability. Further experiments showed that the method could accurately detect the expression of the LKB1 gene, with good linearity observed in the concentration range of 10 pM to 10 nM, and a detection limit as low as 1 pM. Using an arsenic-induced mouse liver injury model, we validated the applicability in complex biological samples and found that this detection system outperformed traditional RT-qPCR in recognizing gene expression differences, offering higher sensitivity and specificity.