TalantaMar 19, 2026

Using enzymes to protect crRNA for better CRISPR/Cas12a detection

CRISPR Gene Editing Weekly Brief ↗PubMed ↗DOI ↗

Updated Mar 20, 2026

Abstract

RI-CRISPR improves detection sensitivity by nearly twentyfold compared to conventional CRISPR/Cas12a.

The CRISPR/Cas12a system faces challenges due to crRNA instability, which affects its diagnostic performance.
RNase inhibitor (RI)-assisted CRISPR (RI-CRISPR) effectively prevents crRNA degradation.
RI-CRISPR enhances the detection performance and anti-interference capability of the CRISPR system.
Clinical validation with 40 MTB samples demonstrates 100% specificity and 96% sensitivity compared to the GeneXpert assay.

Simplified

The CRISPR/Cas12a system enables precise and rapid nucleic acid recognition through the programmable targeting capability of CRISPR RNA (crRNA). However, the intrinsic instability of crRNA limits the robustness and sensitivity of CRISPR-based molecular diagnosis in practical applications. Herein, we present a simple and general enhancement strategy that suppresses RNase-mediated crRNA degradation. This strategy, termed RNase inhibitor (RI)-assisted CRISPR (RI-CRISPR), leverages RI to specifically prevent crRNA degradation, thereby improving its stability and enhancing the detection performance and anti-interference capability of the CRISPR system. Using influenza A virus (IAV) and mycobacterium tuberculosis (MTB) as model targets, RI-CRISPR improves detection sensitivity by nearly twentyfold compared to conventional CRISPR/Cas12a. Clinical validation using 40 MTB samples, combined with recombinase polymerase amplification (RPA), achieves 100% specificity and 96% sensitivity compared with the GeneXpert assay. Overall, this work provides a practical strategy to enhance sensitivity and robustness of CRISPR-based diagnostics, and is expected to promote further biomedical applications of CRISPR technology.

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Using enzymes to protect crRNA for better CRISPR/Cas12a detection

Abstract

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