Ultrasensitive CRISPR/Cas13a-Mediated Photoelectrochemical Biosensors for Specific and Direct Assay of miRNA-21

Sensitive and specific assay of microRNAs (miRNAs) is beneficial to early disease screening. Herein, we for the first time proposed clustered regularly interspaced short palindromic repeats (CRISPR)/Cas13a-mediated photoelectrochemical biosensors for the direct assay of miRNA-21. In this study, compared with traditional nucleic acid-based signal amplification strategies, the CRISPR/Cas13a system can greatly improve the specificity and sensitivity of target determination due to its accurate recognition and high-efficient trans-cleavage capability without complex nucleic acid sequence design. Moreover, compared with the CRISPR/ Cas12a-based biosensing platform, the developed CRISPR/ Cas13a-mediated biosensor can directly detect RNA targets without signal transduction from RNA to DNA, thereby avoiding signal leakage and distortion. Generally, the proposed biosensor reveals excellent analysis capability with a wider linear range from 1 fM to 5 nM and a lower detection limit of 1 fM. Additionally, it also shows satisfactory stability in the detection of human serum samples and cell lysates, manifesting that it has great application prospects in the areas of early disease diagnosis and biomedical research.

Automated summary

In this study, a novel CRISPR/Cas13a-mediated photoelectrochemical biosensor was proposed for the direct assay of miRNA-21, which showed improved specificity and sensitivity compared to traditional nucleic acid-based strategies. The CRISPR/Cas13a system accurately recognized and efficiently cleaved the target, without the need for complex nucleic acid sequence design. This biosensor demonstrated excellent analysis capability and stability, suggesting promising application prospects in early disease diagnosis and biomedical research.