A CRISPR-based approach using dead Cas9-sgRNA to detect SARS-CoV-2
<p dir="ltr">Rapid, highly specific, and robust diagnostic kits to detect viruses and pathogens are needed to control disease spread and transmission globally. Of the many different methods proposed to diagnose COVID-19 infection, CRISPR-based detection of nucleic acids tests are amo...
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2023
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| _version_ | 1864513507641262080 |
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| author | Mustapha Aouida (417652) |
| author2 | Maryam Saifaldeen (12561970) Dana E. Al-Ansari (16327350) Sara Taleb (11264352) Ali Ait Hssain (11264358) Dindial Ramotar (208416) |
| author2_role | author author author author author |
| author_facet | Mustapha Aouida (417652) Maryam Saifaldeen (12561970) Dana E. Al-Ansari (16327350) Sara Taleb (11264352) Ali Ait Hssain (11264358) Dindial Ramotar (208416) |
| author_role | author |
| dc.creator.none.fl_str_mv | Mustapha Aouida (417652) Maryam Saifaldeen (12561970) Dana E. Al-Ansari (16327350) Sara Taleb (11264352) Ali Ait Hssain (11264358) Dindial Ramotar (208416) |
| dc.date.none.fl_str_mv | 2023-06-14T09:00:00Z |
| dc.identifier.none.fl_str_mv | 10.3389/fmolb.2023.1201347 |
| dc.relation.none.fl_str_mv | https://figshare.com/articles/journal_contribution/A_CRISPR-based_approach_using_dead_Cas9-sgRNA_to_detect_SARS-CoV-2/26788474 |
| dc.rights.none.fl_str_mv | CC BY 4.0 info:eu-repo/semantics/openAccess |
| dc.subject.none.fl_str_mv | Biological sciences Genetics Biomedical and clinical sciences Clinical sciences Immunology SARS-CoV-2 M gene dead Cas9 restriction enzymes RT-RPA diagnostics |
| dc.title.none.fl_str_mv | A CRISPR-based approach using dead Cas9-sgRNA to detect SARS-CoV-2 |
| dc.type.none.fl_str_mv | Text Journal contribution info:eu-repo/semantics/publishedVersion text contribution to journal |
| description | <p dir="ltr">Rapid, highly specific, and robust diagnostic kits to detect viruses and pathogens are needed to control disease spread and transmission globally. Of the many different methods proposed to diagnose COVID-19 infection, CRISPR-based detection of nucleic acids tests are among the most prominent. Here, we describe a new way of using CRISPR/Cas systems as a rapid and highly specific tool to detect the SARS-CoV-2 virus using the in vitro dCas9-sgRNA-based technique. As a proof of concept, we used a synthetic DNA of the M gene, one of the SARS-CoV-2 virus genes, and demonstrated that we can specifically inactivate unique restriction enzyme sites on this gene using CRISPR/Cas multiplexing of dCas9-sgRNA-BbsI and dCas9-sgRNA-XbaI. These complexes recognize and bind to the target sequence spanning the BbsI and XbaI restriction enzyme sites, respectively, and protect the M gene from digestion by BbsI and/or XbaI. We further demonstrated that this approach can be used to detect the M gene when expressed in human cells and from individuals infected with SARS-CoV-2. We refer to this approach as dead Cas9 Protects Restriction Enzyme Sites, and believe that it has the potential to be applied as a diagnostic tool for many DNA/RNA pathogens.</p><h2>Other Information</h2><p dir="ltr">Published in: Frontiers in Molecular Biosciences<br>License: <a href="https://creativecommons.org/licenses/by/4.0/" target="_blank">https://creativecommons.org/licenses/by/4.0/</a><br>See article on publisher's website: <a href="https://dx.doi.org/10.3389/fmolb.2023.1201347" target="_blank">https://dx.doi.org/10.3389/fmolb.2023.1201347</a></p> |
| eu_rights_str_mv | openAccess |
| id | Manara2_331b379c1c27b007a162fc524413a159 |
| identifier_str_mv | 10.3389/fmolb.2023.1201347 |
| network_acronym_str | Manara2 |
| network_name_str | Manara2 |
| oai_identifier_str | oai:figshare.com:article/26788474 |
| publishDate | 2023 |
| repository.mail.fl_str_mv | |
| repository.name.fl_str_mv | |
| repository_id_str | |
| rights_invalid_str_mv | CC BY 4.0 |
| spelling | A CRISPR-based approach using dead Cas9-sgRNA to detect SARS-CoV-2Mustapha Aouida (417652)Maryam Saifaldeen (12561970)Dana E. Al-Ansari (16327350)Sara Taleb (11264352)Ali Ait Hssain (11264358)Dindial Ramotar (208416)Biological sciencesGeneticsBiomedical and clinical sciencesClinical sciencesImmunologySARS-CoV-2M genedead Cas9restriction enzymesRT-RPAdiagnostics<p dir="ltr">Rapid, highly specific, and robust diagnostic kits to detect viruses and pathogens are needed to control disease spread and transmission globally. Of the many different methods proposed to diagnose COVID-19 infection, CRISPR-based detection of nucleic acids tests are among the most prominent. Here, we describe a new way of using CRISPR/Cas systems as a rapid and highly specific tool to detect the SARS-CoV-2 virus using the in vitro dCas9-sgRNA-based technique. As a proof of concept, we used a synthetic DNA of the M gene, one of the SARS-CoV-2 virus genes, and demonstrated that we can specifically inactivate unique restriction enzyme sites on this gene using CRISPR/Cas multiplexing of dCas9-sgRNA-BbsI and dCas9-sgRNA-XbaI. These complexes recognize and bind to the target sequence spanning the BbsI and XbaI restriction enzyme sites, respectively, and protect the M gene from digestion by BbsI and/or XbaI. We further demonstrated that this approach can be used to detect the M gene when expressed in human cells and from individuals infected with SARS-CoV-2. We refer to this approach as dead Cas9 Protects Restriction Enzyme Sites, and believe that it has the potential to be applied as a diagnostic tool for many DNA/RNA pathogens.</p><h2>Other Information</h2><p dir="ltr">Published in: Frontiers in Molecular Biosciences<br>License: <a href="https://creativecommons.org/licenses/by/4.0/" target="_blank">https://creativecommons.org/licenses/by/4.0/</a><br>See article on publisher's website: <a href="https://dx.doi.org/10.3389/fmolb.2023.1201347" target="_blank">https://dx.doi.org/10.3389/fmolb.2023.1201347</a></p>2023-06-14T09:00:00ZTextJournal contributioninfo:eu-repo/semantics/publishedVersiontextcontribution to journal10.3389/fmolb.2023.1201347https://figshare.com/articles/journal_contribution/A_CRISPR-based_approach_using_dead_Cas9-sgRNA_to_detect_SARS-CoV-2/26788474CC BY 4.0info:eu-repo/semantics/openAccessoai:figshare.com:article/267884742023-06-14T09:00:00Z |
| spellingShingle | A CRISPR-based approach using dead Cas9-sgRNA to detect SARS-CoV-2 Mustapha Aouida (417652) Biological sciences Genetics Biomedical and clinical sciences Clinical sciences Immunology SARS-CoV-2 M gene dead Cas9 restriction enzymes RT-RPA diagnostics |
| status_str | publishedVersion |
| title | A CRISPR-based approach using dead Cas9-sgRNA to detect SARS-CoV-2 |
| title_full | A CRISPR-based approach using dead Cas9-sgRNA to detect SARS-CoV-2 |
| title_fullStr | A CRISPR-based approach using dead Cas9-sgRNA to detect SARS-CoV-2 |
| title_full_unstemmed | A CRISPR-based approach using dead Cas9-sgRNA to detect SARS-CoV-2 |
| title_short | A CRISPR-based approach using dead Cas9-sgRNA to detect SARS-CoV-2 |
| title_sort | A CRISPR-based approach using dead Cas9-sgRNA to detect SARS-CoV-2 |
| topic | Biological sciences Genetics Biomedical and clinical sciences Clinical sciences Immunology SARS-CoV-2 M gene dead Cas9 restriction enzymes RT-RPA diagnostics |