BRET-based biosensors for SARS-CoV-2 oligonucleotide detection
<p dir="ltr">The need for the early detection of emerging pathogenic viruses and their newer variants has driven the urgent demand for developing point-of-care diagnostic tools. Although nucleic acid-based methods such as reverse transcription-quantitative polymerase chain reaction (...
محفوظ في:
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| مؤلفون آخرون: | , , , |
| منشور في: |
2024
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| _version_ | 1864513510705201152 |
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| author | Asfia Sultana (19170880) |
| author2 | Anupriya M. Geethakumari (17052375) Zeyaul Islam (5867387) Prasanna R. Kolatkar (124118) Kabir H. Biswas (5705864) |
| author2_role | author author author author |
| author_facet | Asfia Sultana (19170880) Anupriya M. Geethakumari (17052375) Zeyaul Islam (5867387) Prasanna R. Kolatkar (124118) Kabir H. Biswas (5705864) |
| author_role | author |
| dc.creator.none.fl_str_mv | Asfia Sultana (19170880) Anupriya M. Geethakumari (17052375) Zeyaul Islam (5867387) Prasanna R. Kolatkar (124118) Kabir H. Biswas (5705864) |
| dc.date.none.fl_str_mv | 2024-06-03T06:00:00Z |
| dc.identifier.none.fl_str_mv | 10.3389/fbioe.2024.1353479 |
| dc.relation.none.fl_str_mv | https://figshare.com/articles/journal_contribution/BRET-based_biosensors_for_SARS-CoV-2_oligonucleotide_detection/26324851 |
| dc.rights.none.fl_str_mv | CC BY 4.0 info:eu-repo/semantics/openAccess |
| dc.subject.none.fl_str_mv | Biomedical and clinical sciences Medical biotechnology Engineering Biomedical engineering biosensor bioluminescence bioluminescence resonance energy transfer COVID-19 SARS-CoV-2 molecular beacon |
| dc.title.none.fl_str_mv | BRET-based biosensors for SARS-CoV-2 oligonucleotide detection |
| dc.type.none.fl_str_mv | Text Journal contribution info:eu-repo/semantics/publishedVersion text contribution to journal |
| description | <p dir="ltr">The need for the early detection of emerging pathogenic viruses and their newer variants has driven the urgent demand for developing point-of-care diagnostic tools. Although nucleic acid-based methods such as reverse transcription-quantitative polymerase chain reaction (RT-qPCR) and loop-mediated isothermal amplification (LAMP) have been developed, a more facile and robust platform is still required. To address this need, as a proof-of-principle study, we engineered a prototype—the versatile, sensitive, rapid, and cost-effective bioluminescence resonance energy transfer (BRET)-based biosensor for oligonucleotide detection (BioOD). Specifically, we designed BioODs against the SARS-CoV-2 parental (Wuhan strain) and B.1.617.2 Delta variant through the conjugation of specific, fluorescently modified molecular beacons (sensor module) through a complementary oligonucleotide handle DNA functionalized with the NanoLuc (NLuc) luciferase protein such that the dissolution of the molecular beacon loop upon the binding of the viral oligonucleotide will result in a decrease in BRET efficiency and, thus, a change in the bioluminescence spectra. Following the assembly of the BioODs, we determined their kinetics response, affinity for variant-specific oligonucleotides, and specificity, and found them to be rapid and highly specific. Furthermore, the decrease in BRET efficiency of the BioODs in the presence of viral oligonucleotides can be detected as a change in color in cell phone camera images. We envisage that the BioODs developed here will find application in detecting viral infections with variant specificity in a point-of-care-testing format, thus aiding in large-scale viral infection surveillance.</p><h2>Other Information</h2><p dir="ltr">Published in: Frontiers in Bioengineering and Biotechnology<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/fbioe.2024.1353479" target="_blank">https://dx.doi.org/10.3389/fbioe.2024.1353479</a></p> |
| eu_rights_str_mv | openAccess |
| id | Manara2_99c81f304ffd3b5c40d62ab90c91baf4 |
| identifier_str_mv | 10.3389/fbioe.2024.1353479 |
| network_acronym_str | Manara2 |
| network_name_str | Manara2 |
| oai_identifier_str | oai:figshare.com:article/26324851 |
| publishDate | 2024 |
| repository.mail.fl_str_mv | |
| repository.name.fl_str_mv | |
| repository_id_str | |
| rights_invalid_str_mv | CC BY 4.0 |
| spelling | BRET-based biosensors for SARS-CoV-2 oligonucleotide detectionAsfia Sultana (19170880)Anupriya M. Geethakumari (17052375)Zeyaul Islam (5867387)Prasanna R. Kolatkar (124118)Kabir H. Biswas (5705864)Biomedical and clinical sciencesMedical biotechnologyEngineeringBiomedical engineeringbiosensorbioluminescencebioluminescence resonance energy transferCOVID-19SARS-CoV-2molecular beacon<p dir="ltr">The need for the early detection of emerging pathogenic viruses and their newer variants has driven the urgent demand for developing point-of-care diagnostic tools. Although nucleic acid-based methods such as reverse transcription-quantitative polymerase chain reaction (RT-qPCR) and loop-mediated isothermal amplification (LAMP) have been developed, a more facile and robust platform is still required. To address this need, as a proof-of-principle study, we engineered a prototype—the versatile, sensitive, rapid, and cost-effective bioluminescence resonance energy transfer (BRET)-based biosensor for oligonucleotide detection (BioOD). Specifically, we designed BioODs against the SARS-CoV-2 parental (Wuhan strain) and B.1.617.2 Delta variant through the conjugation of specific, fluorescently modified molecular beacons (sensor module) through a complementary oligonucleotide handle DNA functionalized with the NanoLuc (NLuc) luciferase protein such that the dissolution of the molecular beacon loop upon the binding of the viral oligonucleotide will result in a decrease in BRET efficiency and, thus, a change in the bioluminescence spectra. Following the assembly of the BioODs, we determined their kinetics response, affinity for variant-specific oligonucleotides, and specificity, and found them to be rapid and highly specific. Furthermore, the decrease in BRET efficiency of the BioODs in the presence of viral oligonucleotides can be detected as a change in color in cell phone camera images. We envisage that the BioODs developed here will find application in detecting viral infections with variant specificity in a point-of-care-testing format, thus aiding in large-scale viral infection surveillance.</p><h2>Other Information</h2><p dir="ltr">Published in: Frontiers in Bioengineering and Biotechnology<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/fbioe.2024.1353479" target="_blank">https://dx.doi.org/10.3389/fbioe.2024.1353479</a></p>2024-06-03T06:00:00ZTextJournal contributioninfo:eu-repo/semantics/publishedVersiontextcontribution to journal10.3389/fbioe.2024.1353479https://figshare.com/articles/journal_contribution/BRET-based_biosensors_for_SARS-CoV-2_oligonucleotide_detection/26324851CC BY 4.0info:eu-repo/semantics/openAccessoai:figshare.com:article/263248512024-06-03T06:00:00Z |
| spellingShingle | BRET-based biosensors for SARS-CoV-2 oligonucleotide detection Asfia Sultana (19170880) Biomedical and clinical sciences Medical biotechnology Engineering Biomedical engineering biosensor bioluminescence bioluminescence resonance energy transfer COVID-19 SARS-CoV-2 molecular beacon |
| status_str | publishedVersion |
| title | BRET-based biosensors for SARS-CoV-2 oligonucleotide detection |
| title_full | BRET-based biosensors for SARS-CoV-2 oligonucleotide detection |
| title_fullStr | BRET-based biosensors for SARS-CoV-2 oligonucleotide detection |
| title_full_unstemmed | BRET-based biosensors for SARS-CoV-2 oligonucleotide detection |
| title_short | BRET-based biosensors for SARS-CoV-2 oligonucleotide detection |
| title_sort | BRET-based biosensors for SARS-CoV-2 oligonucleotide detection |
| topic | Biomedical and clinical sciences Medical biotechnology Engineering Biomedical engineering biosensor bioluminescence bioluminescence resonance energy transfer COVID-19 SARS-CoV-2 molecular beacon |