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Título: | Multiplexable virus detection by CRISPR-Cas9-mediated strand displacement |
Autor: | Márquez-Costa, Rosa; Montagud-Martínez, Roser CSIC; Marqués, M. Carmen ; Heras-Hernández, María CSIC; Albert, Eliseo; Navarro, David; Daròs Arnau, José Antonio CSIC ORCID ; Ruiz, Raúl CSIC ORCID; Rodrigo, Guillermo CSIC ORCID | Palabras clave: | CRISPR diagnostics DNA nanotechnology Synthetic biology |
Fecha de publicación: | 28-nov-2022 | Editor: | MedRxiv | Citación: | MedRxiv: 10.1101/2022.11.23.22282642 (2022) | Resumen: | Recurrent disease outbreaks caused by different viruses, including the novel respiratory virus SARS-CoV-2, are challenging our society at a global scale; so better and handier virus detection methods would enable a faster response. Here, we present a novel nucleic acid detection strategy based on CRISPR-Cas9, whose mode of action relies on strand displacement rather than on collateral catalysis, using the Streptococcus pyogenes Cas9 nuclease. Given a pre-amplification process, a suitable molecular beacon interacts with the ternary CRISPR complex upon targeting to produce a fluorescent signal. We show that SARS-CoV-2 DNA amplicons generated from patient samples can be detected with CRISPR-Cas9. Moreover, we show that CRISPR-Cas9 allows the simultaneous detection of different DNA amplicons with the same nuclease, either to detect different SARS-CoV-2 regions or different respiratory viruses. Collectively, this CRISPR-Cas9 R-loop usage for molecular beacon opening (COLUMBO) platform allows a multiplexed detection in a single tube, complements the existing CRISPR-based methods, and displays diagnostic potential. | Versión del editor: | https://doi.org/10.1101/2022.11.23.22282642 | URI: | http://hdl.handle.net/10261/284005 | DOI: | 10.1101/2022.11.23.22282642 |
Aparece en las colecciones: | (PTI Salud Global) Colección Especial COVID-19 (I2SysBio) Artículos (IBMCP) Artículos |
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