Simultaneous protein and RNA analysis in single extracellular vesicles, including viruses
The individual detection of human immunodeficiency virus (HIV) virions and resolution from extracellular vesicles (EVs) during analysis is a difficult challenge. Infectious enveloped virions and nonviral EVs are released simultaneously by HIV-infected host cells, in addition to hybrid viral EVs cont...
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| Otros Autores: | , , , , , , , |
| Formato: | article |
| Lenguaje: | inglés |
| Publicado: |
2024
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| Acceso en línea: | https://hdl.handle.net/20.500.12008/50874 |
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| _version_ | 1868890121683599360 |
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| author | Troyer, Zach |
| author2 | Gololobova, Olesia Koppula, Aakash Liao, Zhaohao Horns, Felix Elowitz, Michael B Tosar Rovira, Juan Pablo Batish, Mona Witwer, Kenneth |
| author2_role | author author author author author author author author |
| author_browse | Batish, Mona Elowitz, Michael B Gololobova, Olesia Horns, Felix Koppula, Aakash Liao, Zhaohao Tosar Rovira, Juan Pablo Troyer, Zach Witwer, Kenneth |
| author_facet | Troyer, Zach Gololobova, Olesia Koppula, Aakash Liao, Zhaohao Horns, Felix Elowitz, Michael B Tosar Rovira, Juan Pablo Batish, Mona Witwer, Kenneth |
| author_role | author |
| collection | COLIBRI |
| dc.contributor.none.fl_str_mv | Troyer Zach Gololobova Olesia Koppula Aakash Liao Zhaohao Horns Felix Elowitz Michael B Tosar Rovira Juan Pablo, Universidad de la República (Uruguay). Facultad de Ciencias. Centro de Investigaciones Nucleares. Batish Mona Witwer Kenneth |
| dc.creator.none.fl_str_mv | Troyer, Zach Gololobova, Olesia Koppula, Aakash Liao, Zhaohao Horns, Felix Elowitz, Michael B Tosar Rovira, Juan Pablo Batish, Mona Witwer, Kenneth |
| dc.date.none.fl_str_mv | 2024 2025-08-04T17:02:22Z 2025-08-04T17:02:22Z |
| dc.format.none.fl_str_mv | 17 h application/pdf |
| dc.identifier.none.fl_str_mv | Troyer, Z, Gololobova, O, Koppula, A [y otros autores]. "Simultaneous protein and RNA analysis in single extracellular vesicles, including viruses". ACS Nano. [en línea] 2024, 18(39): 26568−26584. 17 h. DOI: 10.1021/acsnano.4c03679 1936-0851 https://hdl.handle.net/20.500.12008/50874 10.1021/acsnano.4c03679 |
| dc.language.none.fl_str_mv | en eng |
| dc.publisher.none.fl_str_mv | ACS Publications |
| dc.relation.none.fl_str_mv | ACS Nano, 2024, 18(39): 26568−26584. |
| dc.rights.none.fl_str_mv | info:eu-repo/semantics/openAccess Licencia Creative Commons Atribución - No Comercial - Sin Derivadas (CC - By-NC-ND 4.0) |
| dc.source.none.fl_str_mv | reponame:COLIBRI instname:Universidad de la República instacron:Universidad de la República |
| dc.subject.none.fl_str_mv | Sngle-particle Single-molecule HIV Extracellular vesicle RNA Protein SP-IRIS |
| dc.title.none.fl_str_mv | Simultaneous protein and RNA analysis in single extracellular vesicles, including viruses |
| dc.type.none.fl_str_mv | Artículo info:eu-repo/semantics/article info:eu-repo/semantics/publishedVersion |
| description | The individual detection of human immunodeficiency virus (HIV) virions and resolution from extracellular vesicles (EVs) during analysis is a difficult challenge. Infectious enveloped virions and nonviral EVs are released simultaneously by HIV-infected host cells, in addition to hybrid viral EVs containing combinations of HIV and host components but lacking replicative ability. Complicating the issue, EVs and enveloped virions are both delimited by a lipid bilayer and share similar size and density. The feature that distinguishes infectious virions from host and hybrid EVs is the HIV genomic RNA (gRNA), which allows the virus to replicate. Singleparticle analysis techniques, which provide snapshots of single biological nanoparticles, could resolve infectious virions from EVs. However, current single-particle analysis techniques focus mainly on protein detection, which fail to resolve hybrid EVs from infectious virions. A method to simultaneously detect viral protein and internal gRNA in the same particle would allow resolution of infectious HIV from EVs and noninfectious virions. Here, we introduce SPIRFISH, a high-throughput method for single-particle protein and RNA analysis, combining single particle interferometric reflectance imaging sensor with singlemolecule fluorescence in situ hybridization. Using SPIRFISH, we detect HIV-1 envelope protein gp120 and genomic RNA within single infectious virions, allowing resolution against EV background and noninfectious virions. We further show that SPIRFISH can be used to detect specific RNAs within EVs. This may have major utility for EV therapeutics, which are increasingly focused on EV-mediated RNA delivery. SPIRFISH should enable single particle analysis of a broad class of RNAcontaining nanoparticles. |
| eu_rights_str_mv | openAccess |
| format | article |
| id | anni_8a2e4ee826ba8ba0e4820ce4c778df0d |
| identifier_str_mv | Troyer, Z, Gololobova, O, Koppula, A [y otros autores]. "Simultaneous protein and RNA analysis in single extracellular vesicles, including viruses". ACS Nano. [en línea] 2024, 18(39): 26568−26584. 17 h. DOI: 10.1021/acsnano.4c03679 1936-0851 10.1021/acsnano.4c03679 |
| instacron_str | Universidad de la República |
| institution | Universidad de la República |
| instname_str | Universidad de la República |
| language | eng |
| language_invalid_str_mv | en |
| network_acronym_str | anni |
| network_name_str | oai-lr-anni |
| oai_identifier_str | oai:colibri.udelar.edu.uy:20.500.12008/50874 |
| publishDate | 2024 |
| publishDateSort | 2024 |
| publisher.none.fl_str_mv | ACS Publications |
| reponame_str | COLIBRI |
| repository.mail.fl_str_mv | |
| repository.name.fl_str_mv | |
| repository_id_str | |
| rights_invalid_str_mv | Licencia Creative Commons Atribución - No Comercial - Sin Derivadas (CC - By-NC-ND 4.0) |
| spelling | Simultaneous protein and RNA analysis in single extracellular vesicles, including virusesTroyer, ZachGololobova, OlesiaKoppula, AakashLiao, ZhaohaoHorns, FelixElowitz, Michael BTosar Rovira, Juan PabloBatish, MonaWitwer, KennethSngle-particleSingle-moleculeHIVExtracellular vesicleRNAProteinSP-IRISThe individual detection of human immunodeficiency virus (HIV) virions and resolution from extracellular vesicles (EVs) during analysis is a difficult challenge. Infectious enveloped virions and nonviral EVs are released simultaneously by HIV-infected host cells, in addition to hybrid viral EVs containing combinations of HIV and host components but lacking replicative ability. Complicating the issue, EVs and enveloped virions are both delimited by a lipid bilayer and share similar size and density. The feature that distinguishes infectious virions from host and hybrid EVs is the HIV genomic RNA (gRNA), which allows the virus to replicate. Singleparticle analysis techniques, which provide snapshots of single biological nanoparticles, could resolve infectious virions from EVs. However, current single-particle analysis techniques focus mainly on protein detection, which fail to resolve hybrid EVs from infectious virions. A method to simultaneously detect viral protein and internal gRNA in the same particle would allow resolution of infectious HIV from EVs and noninfectious virions. Here, we introduce SPIRFISH, a high-throughput method for single-particle protein and RNA analysis, combining single particle interferometric reflectance imaging sensor with singlemolecule fluorescence in situ hybridization. Using SPIRFISH, we detect HIV-1 envelope protein gp120 and genomic RNA within single infectious virions, allowing resolution against EV background and noninfectious virions. We further show that SPIRFISH can be used to detect specific RNAs within EVs. This may have major utility for EV therapeutics, which are increasingly focused on EV-mediated RNA delivery. SPIRFISH should enable single particle analysis of a broad class of RNAcontaining nanoparticles.ACS PublicationsTroyer ZachGololobova OlesiaKoppula AakashLiao ZhaohaoHorns FelixElowitz Michael BTosar Rovira Juan Pablo, Universidad de la República (Uruguay). Facultad de Ciencias. Centro de Investigaciones Nucleares.Batish MonaWitwer Kenneth2025-08-04T17:02:22Z2025-08-04T17:02:22Z2024Artículoinfo:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersion17 happlication/pdfTroyer, Z, Gololobova, O, Koppula, A [y otros autores]. "Simultaneous protein and RNA analysis in single extracellular vesicles, including viruses". ACS Nano. [en línea] 2024, 18(39): 26568−26584. 17 h. DOI: 10.1021/acsnano.4c036791936-0851https://hdl.handle.net/20.500.12008/5087410.1021/acsnano.4c03679reponame:COLIBRIinstname:Universidad de la Repúblicainstacron:Universidad de la RepúblicaenengACS Nano, 2024, 18(39): 26568−26584.Las obras depositadas en el Repositorio se rigen por la Ordenanza de los Derechos de la Propiedad Intelectual de la Universidad de la República.(Res. Nº 91 de C.D.C. de 8/III/1994 – D.O. 7/IV/1994) y por la Ordenanza del Repositorio Abierto de la Universidad de la República (Res. Nº 16 de C.D.C. de 07/10/2014)info:eu-repo/semantics/openAccessLicencia Creative Commons Atribución - No Comercial - Sin Derivadas (CC - By-NC-ND 4.0)oai:colibri.udelar.edu.uy:20.500.12008/508742026-04-14T10:10:53Z |
| spellingShingle | Simultaneous protein and RNA analysis in single extracellular vesicles, including viruses Troyer, Zach Sngle-particle Single-molecule HIV Extracellular vesicle RNA Protein SP-IRIS |
| status_str | publishedVersion |
| title | Simultaneous protein and RNA analysis in single extracellular vesicles, including viruses |
| title_full | Simultaneous protein and RNA analysis in single extracellular vesicles, including viruses |
| title_fullStr | Simultaneous protein and RNA analysis in single extracellular vesicles, including viruses |
| title_full_unstemmed | Simultaneous protein and RNA analysis in single extracellular vesicles, including viruses |
| title_short | Simultaneous protein and RNA analysis in single extracellular vesicles, including viruses |
| title_sort | Simultaneous protein and RNA analysis in single extracellular vesicles, including viruses |
| topic | Sngle-particle Single-molecule HIV Extracellular vesicle RNA Protein SP-IRIS |
| url | https://hdl.handle.net/20.500.12008/50874 |