Evaluation of miR-155 silencing using a molecular beacon in human lung adenocarcinoma cell line
| Autor(a) principal: | |
|---|---|
| Data de Publicação: | 2024 |
| Outros Autores: | , , |
| Tipo de documento: | Artigo |
| Idioma: | eng |
| Título da fonte: | Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) |
| Texto Completo: | http://hdl.handle.net/10362/167748 |
Resumo: | POCI-01-0145-FEDER-022122 research unit PPBI-Portuguese Platform of BioImaging, co-financed by the FEDER through COMPETE 2020, POCI, PORL, and PIDDAC. project PAPILOMA ref. CENTRO-01-0145-FEDER-181235 and \u201CBolsa de Investigação em Oncologia Dr. Rocha Alves do Núcleo Regional do Centro da Liga Portuguesa Contra o Cancro\u201D; This work was supported by EATRIS, the European infrastructure for translational medicine. Publisher Copyright: © 2024 The Author(s) |
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Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) |
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7160 |
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Evaluation of miR-155 silencing using a molecular beacon in human lung adenocarcinoma cell lineA549 cellsGene silencingGold nanoparticlesmiR-155-3pMolecular beaconAnalytical ChemistrySDG 3 - Good Health and Well-beingPOCI-01-0145-FEDER-022122 research unit PPBI-Portuguese Platform of BioImaging, co-financed by the FEDER through COMPETE 2020, POCI, PORL, and PIDDAC. project PAPILOMA ref. CENTRO-01-0145-FEDER-181235 and \u201CBolsa de Investigação em Oncologia Dr. Rocha Alves do Núcleo Regional do Centro da Liga Portuguesa Contra o Cancro\u201D; This work was supported by EATRIS, the European infrastructure for translational medicine. Publisher Copyright: © 2024 The Author(s)Lung cancer (LC) is a leading cause of global cancer-related deaths, highlighting the development of innovative methods for biomarker detection improving the early diagnostics. microRNAs (miRs) alterations are known to be involved in the initiation and progression of human cancers and can act as biomarkers for diagnostics and treatment. Herein, we develop the application of molecular beacon (MB) technology to monitor miR-155-3p expression in human lung adenocarcinoma A549 cells without complementary DNA synthesis, amplification, or expensive reagents. Furthermore, we produced gold nanoparticles (AuNPs) for delivering antisense oligonucleotides into A549 cells to reduce miR-155-3p expression, which was subsequently detectable using the MB. The MB was designed and structural characterized by Förster Resonance Energy Transfer (FRET)-melting, Circular Dichroism (CD), Nuclear magnetic resonance (NMR), and fluorometric experiments, and then the hybridization conditions were optimized for an in vitro approach involving the detection of miR-155-3p in total RNA extracted from A549 cell line. The expression profile of miR-155-3p was obtained by RT-qPCR. The results demonstrated that MB was properly designed and showed efficacy in targeting miR-155-3p. Furthermore, a limit of detection down to nanomolar concentration was achieved and the specificity of the biosensor was proved. Moreover, the self-assembly of ASOs with AuNPs exhibited exceptional target specificity, effectively silencing miR-155-3p. Notably, compared to lipid-based transfection agent, AuNPs displayed superior silencing efficiency. We highlighted the ability of MB to detect changes in the target gene expression after gene silencing. Overall, this innovative approach represents a promising tool for detecting various biomarkers at the same time, with potential applications in clinical settings.UCIBIO - Applied Molecular Biosciences UnitDCV - Departamento de Ciências da VidaRUNAlexandre, DanielaFernandes, Alexandra R.Baptista, Pedro V.Cruz, Carla2024-05-24T01:02:58Z2024-07-012024-07-01T00:00:00Zinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/article10application/pdfhttp://hdl.handle.net/10362/167748eng0039-9140PURE: 91795354https://doi.org/10.1016/j.talanta.2024.126052info:eu-repo/semantics/openAccessreponame:Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)instname:Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informaçãoinstacron:RCAAP2024-07-01T01:34:07Zoai:run.unl.pt:10362/167748Portal AgregadorONGhttps://www.rcaap.pt/oai/openairemluisa.alvim@gmail.comopendoar:71602024-07-01T01:34:07Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) - Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informaçãofalse |
| dc.title.none.fl_str_mv |
Evaluation of miR-155 silencing using a molecular beacon in human lung adenocarcinoma cell line |
| title |
Evaluation of miR-155 silencing using a molecular beacon in human lung adenocarcinoma cell line |
| spellingShingle |
Evaluation of miR-155 silencing using a molecular beacon in human lung adenocarcinoma cell line Alexandre, Daniela A549 cells Gene silencing Gold nanoparticles miR-155-3p Molecular beacon Analytical Chemistry SDG 3 - Good Health and Well-being |
| title_short |
Evaluation of miR-155 silencing using a molecular beacon in human lung adenocarcinoma cell line |
| title_full |
Evaluation of miR-155 silencing using a molecular beacon in human lung adenocarcinoma cell line |
| title_fullStr |
Evaluation of miR-155 silencing using a molecular beacon in human lung adenocarcinoma cell line |
| title_full_unstemmed |
Evaluation of miR-155 silencing using a molecular beacon in human lung adenocarcinoma cell line |
| title_sort |
Evaluation of miR-155 silencing using a molecular beacon in human lung adenocarcinoma cell line |
| author |
Alexandre, Daniela |
| author_facet |
Alexandre, Daniela Fernandes, Alexandra R. Baptista, Pedro V. Cruz, Carla |
| author_role |
author |
| author2 |
Fernandes, Alexandra R. Baptista, Pedro V. Cruz, Carla |
| author2_role |
author author author |
| dc.contributor.none.fl_str_mv |
UCIBIO - Applied Molecular Biosciences Unit DCV - Departamento de Ciências da Vida RUN |
| dc.contributor.author.fl_str_mv |
Alexandre, Daniela Fernandes, Alexandra R. Baptista, Pedro V. Cruz, Carla |
| dc.subject.por.fl_str_mv |
A549 cells Gene silencing Gold nanoparticles miR-155-3p Molecular beacon Analytical Chemistry SDG 3 - Good Health and Well-being |
| topic |
A549 cells Gene silencing Gold nanoparticles miR-155-3p Molecular beacon Analytical Chemistry SDG 3 - Good Health and Well-being |
| description |
POCI-01-0145-FEDER-022122 research unit PPBI-Portuguese Platform of BioImaging, co-financed by the FEDER through COMPETE 2020, POCI, PORL, and PIDDAC. project PAPILOMA ref. CENTRO-01-0145-FEDER-181235 and \u201CBolsa de Investigação em Oncologia Dr. Rocha Alves do Núcleo Regional do Centro da Liga Portuguesa Contra o Cancro\u201D; This work was supported by EATRIS, the European infrastructure for translational medicine. Publisher Copyright: © 2024 The Author(s) |
| publishDate |
2024 |
| dc.date.none.fl_str_mv |
2024-05-24T01:02:58Z 2024-07-01 2024-07-01T00:00:00Z |
| dc.type.status.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
| dc.type.driver.fl_str_mv |
info:eu-repo/semantics/article |
| format |
article |
| status_str |
publishedVersion |
| dc.identifier.uri.fl_str_mv |
http://hdl.handle.net/10362/167748 |
| url |
http://hdl.handle.net/10362/167748 |
| dc.language.iso.fl_str_mv |
eng |
| language |
eng |
| dc.relation.none.fl_str_mv |
0039-9140 PURE: 91795354 https://doi.org/10.1016/j.talanta.2024.126052 |
| dc.rights.driver.fl_str_mv |
info:eu-repo/semantics/openAccess |
| eu_rights_str_mv |
openAccess |
| dc.format.none.fl_str_mv |
10 application/pdf |
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reponame:Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) instname:Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação instacron:RCAAP |
| instname_str |
Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação |
| instacron_str |
RCAAP |
| institution |
RCAAP |
| reponame_str |
Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) |
| collection |
Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) |
| repository.name.fl_str_mv |
Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) - Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação |
| repository.mail.fl_str_mv |
mluisa.alvim@gmail.com |
| _version_ |
1817546016976535552 |