Rapid, inexpensive, and sensitive nucleic acid detection may aid point-of-care pathogen detection, genotyping, and disease monitoring. We combine the RNA- targeting CRISPR effector Cas13 with isothermal amplification to establish a CRISPR-based diagnostic (CRISPR-Dx), providing rapid DNA or RNA detection with attomolar sensitivity and single-base mismatch specificity. We use this Cas13a-based molecular detection platform, termed SHERLOCK, to detect specific strains of Zika and Dengue virus, distinguish pathogenic bacteria, genotype human DNA, and identify cell-free tumor DNA mutations. By leveraging CRISPR enzymology and lateral flow devices, we demonstrate further improvements on SHERLOCK, including multiplexing and instrument-free readout. CRISPR-Dx forms the basis for multiplexable, portable, rapid, and quantitative detection of nucleic acids.
1. At this seminar, attendees will gain and understanding of CRISPR enzyme diversity and the distinguish characteristics of Cas12 and Cas13 enzymes.
2. After attending this seminar, attendees will be able to describe the development of CRISPR tools for diagnostics in the field and the clinic