The CRISPR-CS platform enabled unamplified nucleic acid electrochemical assay in <5 minutes with a detection limit of 9.5×10^-20 M and detected targets with up to 98.1% accuracy.
Does a CRISPR/Cas cooperative shearing system improve detection limit and accuracy compared to traditional CRISPR/Cas systems in molecular diagnostics?
The CRISPR-CS system significantly improves the detection limit and accuracy of nucleic acid electrochemical assays for clinical diagnostics.
Abstract The CRISPR/Cas system is a powerful genome editing tool and possesses widespread applications in molecular diagnostics, therapeutics and genetic engineering. But easy folding of the target sequences causes remarkable deterioration of the recognition and shear efficiency in the case of single Cas‐CRISPR RNA (crRNA) duplex. Here, we develop a CRISPR/Cas cooperative shearing (CRISPR‐CS) system. Compared with traditional CRISPR/Cas system, two CRISPR/Cas‐crRNA duplexes simultaneously recognize different sites in the target sequence, increasing recognition possibility and shearing efficiency. Cooperative shearing cuts more methylene blue‐ssDNA reporters on the electrode, enabling unamplified nucleic acid electrochemical assay in less than 5 minutes with a detection limit of 9.5×10 −20 M, 2 to 9 orders of magnitude lower than those of other electrochemical assays. The CRISPR‐CS platform detects monkeypox, human papilloma virus and amyotrophic lateral sclerosis with an accuracy up to 98.1 %, demonstrating the potential application of the efficient cooperative shearing.
Zhao et al. (2024) studied Monkeypox, human papilloma virus, and amyotrophic lateral sclerosis. CRISPR/Cas cooperative shearing (CRISPR-CS) system vs. Traditional CRISPR/Cas system was evaluated on Detection limit and accuracy. The CRISPR-CS platform enabled unamplified nucleic acid electrochemical assay in <5 minutes with a detection limit of 9.5×10^-20 M and detected targets with up to 98.1% accuracy.