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dc.contributor.authorSantiago-Frangos, Andrew
dc.contributor.authorHall, Laina N.
dc.contributor.authorNemudraia, Anna
dc.contributor.authorNemudryi, Artem
dc.contributor.authorKrishna, Pushya
dc.contributor.authorWiegand, Tanner
dc.contributor.authorWilkinson, Royce A.
dc.contributor.authorSnyder, Deann T.
dc.contributor.authorHedges, Jodi F.
dc.contributor.authorCicha, Calvin
dc.contributor.authorLee, Helen H.
dc.contributor.authorGraham, Ava
dc.contributor.authorJutila, Mark A.
dc.contributor.authorTaylor, Matthew P.
dc.contributor.authorWiedenheft, Blake
dc.date.accessioned2022-08-30T22:52:24Z
dc.date.available2022-08-30T22:52:24Z
dc.date.issued2021-06
dc.identifier.citationSantiago-Frangos, A., Hall, L. N., Nemudraia, A., Nemudryi, A., Krishna, P., Wiegand, T., ... & Wiedenheft, B. (2021). Intrinsic signal amplification by type III CRISPR-Cas systems provides a sequence-specific SARS-CoV-2 diagnostic. Cell Reports Medicine, 2(6), 100319.en_US
dc.identifier.issn2666-3791
dc.identifier.urihttps://scholarworks.montana.edu/xmlui/handle/1/17034
dc.description.abstractThere is an urgent need for inexpensive new technologies that enable fast, reliable, and scalable detection of viruses. Here, we repurpose the type III CRISPR-Cas system for sensitive and sequence-specific detection of SARS-CoV-2. RNA recognition by the type III CRISPR complex triggers Cas10-mediated polymerase activity, which simultaneously generates pyrophosphates, protons, and cyclic oligonucleotides. We show that all three Cas10-polymerase products are detectable using colorimetric or fluorometric readouts. We design ten guide RNAs that target conserved regions of SARS-CoV-2 genomes. Multiplexing improves the sensitivity of amplification-free RNA detection from 107 copies/μL for a single guide RNA to 106 copies/μL for ten guides. To decrease the limit of detection to levels that are clinically relevant, we developed a two-pot reaction consisting of RT-LAMP followed by T7-transcription and type III CRISPR-based detection. The two-pot reaction has a sensitivity of 200 copies/μL and is completed using patient samples in less than 30 min.en_US
dc.language.isoen_USen_US
dc.publisherElsevier BVen_US
dc.rightscc-byen_US
dc.rights.urihttps://creativecommons.org/licenses/by/4.0/en_US
dc.subjectsars cov 2en_US
dc.titleIntrinsic signal amplification by type III CRISPR-Cas systems provides a sequence-specific SARS-CoV-2 diagnosticen_US
dc.typeArticleen_US
mus.citation.extentfirstpage1en_US
mus.citation.extentlastpage14en_US
mus.citation.issue6en_US
mus.citation.journaltitleCell Reports Medicineen_US
mus.citation.volume2en_US
mus.identifier.doi10.1016/j.xcrm.2021.100319en_US
mus.relation.collegeCollege of Agricultureen_US
mus.relation.departmentMicrobiology & Cell Biology.en_US
mus.relation.universityMontana State University - Bozemanen_US
mus.data.thumbpage5en_US


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