Multiplexable and Biocomputational Virus Detection by CRISPR-Cas9-Mediated Strand Displacement.
Anal Chem
; 95(25): 9564-9574, 2023 06 27.
Article
in English
| MEDLINE | ID: covidwho-2324755
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 versatile virus detection methods would enable a calculated and 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 preamplification 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. We also 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. Furthermore, we demonstrate that engineered DNA logic circuits can process different SARS-CoV-2 signals detected by the CRISPR complexes. Collectively, this CRISPR-Cas9 R-loop usage for the molecular beacon opening (COLUMBO) platform allows a multiplexed detection in a single tube, complements the existing CRISPR-based methods, and displays diagnostic and biocomputing potential.
Full text:
Available
Collection:
International databases
Database:
MEDLINE
Main subject:
CRISPR-Cas Systems
/
COVID-19
Type of study:
Diagnostic study
Limits:
Humans
Language:
English
Journal:
Anal Chem
Year:
2023
Document Type:
Article
Affiliation country:
Acs.analchem.3c01041
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