Photo-switchable CRISPR electrochemical system enables de-interferential biosensing signal output.

Most clustered regularly interspaced short palindromic repeats (CRISPR)-based electrochemical biosensors lack controllable switching function and are vulnerable to false signals induced by intrinsic amplification signal crosstalk. Thus, exploring strategies that mitigate intrinsic amplification crosstalk with a precise switch would be extremely useful for reliable and accurate bioanalysis. Herein, we develop a photo-switchable CRISPR/Cas12a electrochemical (PSCE) system by introducing a photocleavable (PC) linker and adopting a light-responsive strategy, which enables output of de-interference electrochemical signals and achieves highly sensitive and accurate detection of gene mutations for non-small cell lung cancer (NSCLC). The system decouples nucleic acid amplification (NAA) from signal transduction and produces unique photo-switchable response signals by light irradiation to activate Cas12a activity. This architectural design further separates signal readout from sample pretreatment, suppressing intrinsic amplification-derived signal crosstalk rather than global biological interference to deliver an ultralow limit of detection (LoD). The PSCE achieves 98.1% sensitivity, 92.7% specificity, and 98.9% overall accuracy when tested with 67 clinical samples. Moreover, the exploration of PSCE system in flexible wearable electronics and machine-learning analysis of clinical patient samples demonstrates significant application potential clinical diagnosis of mutation-associated diseases.