CRISPR Enzyme Kinetics for Molecular Diagnostics

CRISPR-diagnostic assays have gained significant interest in the last few years. This has grown rapidly during current COVID-19 pandemic, where CRISPR-diagnostics been frontline contenders for rapid testing solutions. surge research prompts following question: what exactly are achievable limits of detection and associated assay times enabled by kinetics enzymes such as Cas12 Cas13? To explore this question, we here present a model based on Michaelis–Menten enzyme theory applied to CRISPR enzymes. We use develop analytical solutions reaction back-of-the-envelope criteria validate check consistency reported kinetic parameters. our analyses all studies known us, which report Michaelis–Menten-type data CRISPR-associated These include subtypes Cas13 orthologs. found but one study clearly violate at least two three rules therefore that basic physical limits. performed an experimental LbCas12a with both ssDNA dsDNA activators these its predicted scaling. The validated is used time scales degree completion practically relevant target concentrations applicable assays. results broad implications emerging, amplification-free CRISPR-detection methods.