CRISPR-dCas13a-driven electrochemical biosensor for label-free detection of epilepsy-associated MicroRNAs

Abstract

Early detection and monitoring of epileptic seizures are crucial for timely intervention and effective management. Conventional diagnostic techniques, however, are often associated with high costs and operational complexity, underscoring the urgent need for innovative, accessible alternatives. This study presents an analytical proof-of-concept for a CRISPR-based electrochemical biosensor that enables label-free detection of epilepsy-associated microRNA biomarkers under controlled experimental conditions. We developed a label-free sensing platform by integrating a catalytically inactive Cas13a (dCas13a) complex with synthetic guide RNAs (sgRNAs) on a chemically modified screen-printed gold electrode (SPGE). This design enables highly specific binding and quantification of target microRNAs without cleavage. The platform specifically targets miR-143-3p and miR-145-5p, which are recognized as key biomarkers due to their significant modulation during epileptic seizures and their correlation with seizure duration. Sensitive detection of these microRNAs may provide supportive molecular information on seizure-associated biological changes and could contribute to future studies investigating seizure burden and disease progression. By combining CRISPR technology with electrochemical sensing, this work establishes a simple, amplification-free, and potentially portable biosensing strategy. The platform is intended as a complementary molecular tool that may support future point-of-care applications in epilepsy diagnosis.