Nucleic acid

Electronic engineering

Engineering

Sensitivity (control systems)

Base pair

Biology

Gene

Genotyping

CRISPR

Genotype

Computational biology

Genetics

Genome biology

CRISPR-based nucleic acid detection methods are reported to facilitate rapid and sensitive DNA detection. However, precise DNA detection at the single-base resolution and its wide applications including high-fidelity SNP genotyping remain to be explored. Here we develop a Cas12b-mediated DNA detection (CDetection) strategy, which shows higher sensitivity on examined targets compared with the previously reported Cas12a-based detection platform. Moreover, we show that CDetection can distinguish differences at the single-base level upon combining the optimized tuned guide RNA (tgRNA). Therefore, our findings highlight the high sensitivity and accuracy of CDetection, which provides an efficient and highly practical platform for DNA detection.

CDetection: CRISPR-Cas12b-based DNA detection with sub-attomolar sensitivity and single-base specificity