TL;DR
This cost-effective, adaptable strategy can streamline RPA-CRISPR diagnostics for infectious disease surveillance and convert established two-step assays for Burkholderia pseudomallei and Brucella into one-pot formats.
This work presents a broadly applicable one-pot detection strategy termed mutant scaffold-mediated RPA-CRISPR/Cas12a (MS-CRISPR), and introduces rationally designed point mutations into the crRNA scaffold to moderately attenuate Cas12a activation and cleavage kinetics.
CATNAP (Cas trans -nuclease detection of amplified products), a method that integrates isothermal linear DNA amplification with Cas12a detection in a single reaction, bridges the sensitivity gap in CRISPR diagnostics while maintaining simplicity, making accurate disease detection more accessible in resource-limited setti...
A split-crRNA-based CRISPR/Cas12a assay integrated with a lateral flow assay is developed for rapid, sensitive detection of bacterial 16S rRNA within 1 h, without pre-amplification, presenting a promising approach for effective food safety monitoring and prevention of infectious diseases.
A crRNA-universal, sensitive and specific CRISPR-Cas12a detection platform, termed DESIC (double-end blocker and split-input mediated CRISPR-Cas12a system), for single-base mutation detection, targeting four prevalent pancreatic cancer KRAS mutations and optimized the system to achieve optimal discrimination.
The strict requirement for a canonical TTTV protospacer adjacent motif (PAM) restricts target-site selection in clustered regularly interspaced short palindromic repeats/CRISPR-associated protein 12a (CRISPR/Cas12a)-based diagnostics and limits the flexibility of assay design. Whether suboptimal PAMs can be effectively...
ABSTRACT Detection of gene mutations related to acute myeloid leukemia (AML) is crucial for its early diagnosis, personalized treatment, and recurrence monitoring. However, accurate detection of diverse mutations occurring at the same locus remains technically challenging. The CRISPR/Cas system, with its unique program...
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