Skip to main content
. 2023 Apr 23;25(7):428–437. doi: 10.1016/j.jmoldx.2023.03.009

Figure 1.

Figure 1

Thermostable Cas enzyme discovery and characterization. A: High-throughput novel Cas12 enzyme discovery pipeline. B: Melting temperature (Tm), as determined by thermal shift assay for 14 novel Cas12a/b enzymes, Leptotrichia wadei LwaCas13 and AacCas12. The horizontal line indicates the preferred Tm of 60°C. C: Real-time SLK assay run at 60°C, testing five novel Cas12a/b enzymes and AacCas12b. The baseline-subtracted RFU signal (relative fluorescence units) is a measure of collateral cleavage activity of each enzyme. SARS-CoV-2 genomic RNA at 200 cp/μL or 10 ng/μL human genomic RNA was used as target, and water, as the no-template control (NTC). Data are expressed as means ± SEM. n = 3 per sample. crRNA, guide RNA; Pfam, database of protein families.