CRISPR-based platform for carbapenemases and emerging viruses detection using Cas12a (Cpf1) effector nuclease

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dc.citation.titleEmerging Microbes & Infections
dc.citation.volume9
dc.creatorCurti, Lucía Ana
dc.creatorPereyra Bonnet, Federico
dc.creatorRepizo, Guillermo Daniel
dc.creatorFay, Jessica Vannina
dc.creatorSalvatierra, Karina
dc.creatorBlariza, María José
dc.creatorIbáñez Alegre, Macarena
dc.creatorRinflerch, Adriana Raquel
dc.creatorMiretti, Marcos
dc.creatorGiménez, Carla Alejandra
dc.date.accessioned2024-07-05T14:54:10Z
dc.date.available2024-07-05T14:54:10Z
dc.date.issued2020-06-02
dc.description.abstractCRISPR-Cas12a (also called Cpf1) has been commonly used for genomic editing, based on its ability to generate precise double-stranded DNA (dsDNA) breaks. Recently, it was demonstrated that Cas12a exhibits unspecific ssDNAse activity upon target recognition. This feature allows CRISPR-Cas to be coupled with a ssDNA reporter and generate a fast, accurate and ultrasensitive molecular detection method. Here, we demonstrate that Cas12a was able to detect DNA target sequences corresponding to carbapenemases resistance genes such as KPC, NDM and OXA. Also, with the addition of a reverse-transcription step, we were able to detect viral RNA sequences from DENV, ZIKV and HANTV genomes. In all cases, assay run time was less than two hours. Additionally, we report attomolar levels of detection. This methodology was validated using clinical samples from patients infected with Dengue virus. Reactions were visualized by detection of a fluorescent signal, as well as by the use of a simple lateral flow strip. These results indicate that Cas12a is able to detect both DNA and RNA targets, making it an appropriate and convenient tool to detect all types of pathogens.
dc.description.filFil: Curti, Lucía Ana. Universidad de Buenos Aires. Instituto de Investigaciones en Producción Animal (INPA-CONICET); Argentina.
dc.description.filFil: Pereyra Bonnet, Federico. Universidad de Buenos Aires. Instituto de Investigaciones en Producción Animal (INPA-CONICET); Argentina.
dc.description.filFil: Pereyra Bonnet, Federico. CASPR Biotech; United States.
dc.description.filFil: Repizo, Guillermo Daniel. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Instituto de Biología Molecular y Celular de Rosario (IBR-CONICET); Argentina.
dc.description.filFil: Fay, Jessica Vannina. Universidad Nacional de Misiones. Facultad de Ciencias Exactas, Químicas y Naturales. Instituto de Biología Subtropica. Laboratorio GIGA; Argentina.
dc.description.filFil: Salvatierra, Karina. Universidad Nacional de Misiones. Facultad de Ciencias Exactas, Químicas y Naturales. Instituto de Biología Subtropica. Laboratorio GIGA; Argentina.
dc.description.filFil: Blariza, María José. Universidad Nacional de Misiones. Facultad de Ciencias Exactas, Químicas y Naturales. Instituto de Biología Subtropica. Laboratorio GIGA; Argentina.
dc.description.filFil: Ibáñez Alegre, Macarena. Universidad Nacional de Misiones. Facultad de Ciencias Exactas, Químicas y Naturales. Instituto de Biología Subtropica. Laboratorio GIGA; Argentina.
dc.description.filFil: Rinflerch, Adriana Raquel. Universidad Nacional de Misiones. Facultad de Ciencias Exactas, Químicas y Naturales. Instituto de Biología Subtropica. Laboratorio GIGA; Argentina.
dc.description.filFil: Miretti, Marcos. Universidad Nacional de Misiones. Facultad de Ciencias Exactas, Químicas y Naturales. Instituto de Biología Subtropica. Laboratorio GIGA; Argentina.
dc.description.filFil: Giménez, Carla Alejandra. CASPR Biotech; United States.
dc.description.sponsorshipCaspr Biotech
dc.description.sponsorshipConsejo Nacional de Investigaciones Científicas y Técnicas (CONICET)
dc.description.versionpeerreviewed
dc.format.extent1-9
dc.identifier.issn2222-1751
dc.identifier.urihttps://hdl.handle.net/2133/27437
dc.language.isoen
dc.publisherTaylor & Francis
dc.relation.publisherversionhttps://doi.org/10.1080/22221751.2020.1763857
dc.relation.publisherversionhttps://www.tandfonline.com/doi/full/10.1080/22221751.2020.1763857
dc.rightsopenAccess
dc.rights.holderCurti, Lucía Ana
dc.rights.holderPereyra Bonnet, Federico
dc.rights.holderRepizo, Guillermo Daniel
dc.rights.holderFay, Jessica Vannina
dc.rights.holderSalvatierra, Karina
dc.rights.holderBlariza, María José
dc.rights.holderIbáñez Alegre, Macarena
dc.rights.holderRinflerch, Adriana Raquel
dc.rights.holderMiretti, Marcos
dc.rights.holderGiménez, Carla Alejandra
dc.rights.holderUniversidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas
dc.rights.textAttribution 4.0 Internationalen
dc.rights.urihttp://creativecommons.org/licenses/by/4.0/
dc.subjectInfectious diseases
dc.subjectMolecular diagnostic
dc.subjectRISPR/Cas12a
dc.subjectRNA viruses
dc.subjectAntibiotic resistance
dc.titleCRISPR-based platform for carbapenemases and emerging viruses detection using Cas12a (Cpf1) effector nuclease
dc.typearticulo
dc.type.versionpublishedVersion

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(FBIOyF) Departamento de Microbiología - Artículos