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Multiplexable virus detection by CRISPR-Cas9-mediated strand displacement

Marquez-Costa, R.; Montagud-Martinez, R.; Marques, M.-C.; Heras-Hernandez, M.; Albert, E.; Navarro, D.; Daros, J.-A.; Ruiz, R.; Rodrigo, G.

2022-11-28 infectious diseases
10.1101/2022.11.23.22282642 medRxiv
Show abstract

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.

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The top 9 journals account for 50% of the predicted probability mass.

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"Similar papers" are the closest papers from that journal in the model's embedding space. They show what the match is built on, but the ranking comes mostly from a classifier over the whole training set, not from these examples alone.