The application of CRISPR-Cas for single species identification from environmental DNA

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dc.contributor.authorWilliams, Molly-Ann
dc.contributor.authorO'Grady, Joyce
dc.contributor.authorBall, Bernard
dc.contributor.authorCarlsson, Jens
dc.contributor.authorde Eyto, Elvira
dc.contributor.authorMcGinnity, Philip
dc.contributor.authorJennings, Eleanor
dc.contributor.authorRegan, Fiona
dc.contributor.authorParle-McDermott, Anne
dc.contributor.funderMarine Instituteen
dc.date.accessioned2019-06-13T09:38:16Z
dc.date.available2019-06-13T09:38:16Z
dc.date.issued2019-06-08
dc.date.updated2019-06-13T08:22:53Z
dc.description.abstractWe report the first application of CRISPR‐Cas technology to single species detection from environmental DNA (eDNA). Organisms shed and excrete DNA into their environment such as in skin cells and faeces, referred to as environmental DNA (eDNA). Utilising eDNA allows non‐invasive monitoring with increased specificity and sensitivity. Current methods primarily employ PCR‐based techniques to detect a given species from eDNA samples, posing a logistical challenge for on‐site monitoring and potential adaptation to biosensor devices. We have developed an alternative method; coupling isothermal amplification to a CRISPR‐Cas12a detection system. This utilises the collateral cleavage activity of Cas12a, a ribonuclease guided by a highly specific single CRISPR RNA. We used the target species Salmo salar as a proof‐of‐concept test of the specificity of the assay among closely related species and to show the assay is successful at a single temperature of 37°C with signal detection at 535 nM. The specific assay, detects at attomolar sensitivity with rapid detection rates (<2.5 h). This approach simplifies the challenge of building a biosensor device for rapid target species detection in the field and can be easily adapted to detect any species from eDNA samples from a variety of sources enhancing the capabilities of eDNA as a tool for monitoring biodiversity.en
dc.description.sponsorshipMarine Institute (Burrishoole Ecosystem Observatory Network 2020: BEYOND 2020 PBA/FS/16/02)en
dc.description.statusPeer revieweden
dc.description.versionAccepted Versionen
dc.format.mimetypeapplication/pdfen
dc.identifier.citationWilliams, M.-A., O'Grady, J., Ball, B., Carlsson, J., de Eyto, E., McGinnity, P., Jennings, E., Regan, F. and Parle-McDermott, A. (2019) 'The application of CRISPR-Cas for single species identification from environmental DNA', Molecular Ecology Resources. doi: 10.1111/1755-0998.13045en
dc.identifier.doi10.1111/1755-0998.13045en
dc.identifier.eissn1755-0998
dc.identifier.issn1755-098X
dc.identifier.journaltitleMolecular Ecology Resourcesen
dc.identifier.urihttps://hdl.handle.net/10468/8053
dc.language.isoenen
dc.publisherJohn Wiley & Sons, Inc.en
dc.relation.urihttps://onlinelibrary.wiley.com/doi/abs/10.1111/1755-0998.13045
dc.rights© 2019, John Wiley & Sons Inc. This is the peer reviewed version of the following article: Williams, M.-A., O'Grady, J., Ball, B., Carlsson, J., de Eyto, E., McGinnity, P., Jennings, E., Regan, F. and Parle-McDermott, A. (2019) 'The application of CRISPR-Cas for single species identification from environmental DNA', Molecular Ecology Resources. doi: 10.1111/1755-0998.13045, which has been published in final form at https://doi.org/10.1111/1755-0998.13045. This article may be used for non-commercial purposes in accordance with Wiley Terms and Conditions for Use of Self-Archived Versions.en
dc.subjectBiosensoren
dc.subjecteDNAen
dc.subjectEnvironmentalen
dc.subjectFreshwateren
dc.subjectSalmonen
dc.subjectCRISPR-Casen
dc.titleThe application of CRISPR-Cas for single species identification from environmental DNAen
dc.typeArticle (peer-reviewed)en
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