A streamlined proof of concept CRISPR-based environmental biosurveillance platform for the detection of closely related invasive rats
Duran-Vinet, B.; Piaggio, A. J.; Foster, N.; Clark, A.; Salyer, M.; Jeunen, G.-J.; Treece, J.; Ferreira, S.; Collins, C.; Buckelew, S.; Gemmell, N.
Show abstract
Environmental biosurveillance requires technologies that can rapidly translate genomic information into sensitive, specific, and deployable detection tools. Here, we present the first proof-of-concept application of SENTINEL (Smart Environmental Nucleic-acid Tracking using Inference from Neural-networks for Early-warning Localization), integrating artificial-intelligence-guided target discovery, isothermal amplification, CRISPR-based detection, and environmental DNA. Using Rattus exulans, R. norvegicus, and R. rattus as a challenging model of closely related invasive vertebrates, SENTINEL achieved species-specific discrimination without detectable cross-reactivity and sensitivities of 10 aM for plasmid DNA and 10 fg/uL for genomic DNA. Selected assays also showed reproducible semi-quantitative performance and successful eDNA validation. This first application of CRISPR-based environmental biosurveillance to terrestrial invasive vertebrates demonstrates a transferable framework for converting sequence data into field-deployable assays, with potential applications across biosecurity, conservation, agriculture, and environmental monitoring.
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