Production of Potyvirus-Derived Nanoparticles Decorated with a Nanobody in Biofactory Plants
[EN] Viral nanoparticles (VNPs) have recently attracted attention for their use as building blocks for novel materials to support a range of functions of potential interest in nanotechnology and medicine. Viral capsids are ideal for presenting small epitopes by inserting them at an appropriate site...
| Autores: | , , , |
|---|---|
| Tipo de recurso: | artículo |
| Fecha de publicación: | 2022 |
| País: | España |
| Institución: | Universitat Politècnica de València (UPV) |
| Repositorio: | RiuNet. Repositorio Institucional de la Universitat Politécnica de Valéncia |
| Idioma: | inglés |
| OAI Identifier: | oai:riunet.upv.es:10251/193226 |
| Acceso en línea: | https://riunet.upv.es/handle/10251/193226 |
| Access Level: | acceso abierto |
| Palabra clave: | Viral nanoparticle VNP Plant virus Nanobody Potyvirus Zucchini yellow mosaic virus Tobacco etch virus |
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Production of Potyvirus-Derived Nanoparticles Decorated with a Nanobody in Biofactory PlantsMartí, Mari CarmenMerwaiss, FernandoButkovic, AnamarijaDaròs, José-Antonio|||0000-0002-6535-2889Viral nanoparticleVNPPlant virusNanobodyPotyvirusZucchini yellow mosaic virusTobacco etch virus[EN] Viral nanoparticles (VNPs) have recently attracted attention for their use as building blocks for novel materials to support a range of functions of potential interest in nanotechnology and medicine. Viral capsids are ideal for presenting small epitopes by inserting them at an appropriate site on the selected coat protein (CP). VNPs presenting antibodies on their surfaces are considered highly promising tools for therapeutic and diagnostic purposes. Due to their size, nanobodies are an interesting alternative to classic antibodies for surface presentation. Nanobodies are the variable domains of heavy-chain (VHH) antibodies from animals belonging to the family Camelidae, which have several properties that make them attractive therapeutic molecules, such as their small size, simple structure, and high affinity and specificity. In this work, we have produced genetically encoded VNPs derived from two different potyviruses¿the largest group of RNA viruses that infect plants¿decorated with nanobodies. We have created a VNP derived from zucchini yellow mosaic virus (ZYMV) decorated with a nanobody against the green fluorescent protein (GFP) in zucchini (Cucurbita pepo) plants. As reported for other viruses, the expression of ZYMV-derived VNPs decorated with this nanobody was only made possible by including a picornavirus 2A splicing peptide between the fused proteins, which resulted in a mixed population of unmodified and decorated CPs. We have also produced tobacco etch virus (TEV)-derived VNPs in Nicotiana benthamiana plants decorated with the same nanobody against GFP. Strikingly, in this case, VNPs could be assembled by direct fusion of the nanobody to the viral CP with no 2A splicing involved, likely resulting in fully decorated VNPs. For both expression systems, correct assembly and purification of the recombinant VNPs was confirmed by transmission electron microscope; the functionality of the CP-fused nanobody was assessed by western blot and binding assays. In sum, here we report the production of genetically encoded plant-derived VNPs decorated with a nanobody. This system may be an attractive alternative for the sustainable production in plants of nanobody-containing nanomaterials for diagnostic and therapeutic purposes.This research was supported by grant PID 2020-114691RB-I00 from the Ministerio de Ciencia e Innovacion (Spain; co-financed by the European Region Development Fund) through the Agencia Estatal de Investigacion. MM is the recipient of a predoctoral contract (FPU16/05294) from the Ministerio de Educacion, Cultura y Deporte (Spain).Frontiers Media S.A.Instituto Universitario Mixto de Biología Molecular y Celular de PlantasEuropean Regional Development FundMinisterio de Ciencia e InnovaciónMinisterio de Educación, Cultura y DeporteRepositorio Institucional de la Universitat Politècnica de València Riunet20222022-03-31journal articlehttp://purl.org/coar/resource_type/c_6501VoRhttp://purl.org/coar/version/c_970fb48d4fbd8a85info:eu-repo/semantics/articleapplication/pdfhttps://riunet.upv.es/handle/10251/193226reponame:RiuNet. Repositorio Institucional de la Universitat Politécnica de Valénciainstname:Universitat Politècnica de València (UPV)InglésengMinisterio de Educación y Cultura http://dx.doi.org/10.13039/501100003176 FPU16%2F05294 FPU16%2F05294Ministerio de Ciencia e Innovación http://dx.doi.org/10.13039/501100004837 PID2020-114691RB-I00 Biotecnología de virus de plantas: vectores virales y estrategias de resistenciaopen accesshttp://purl.org/coar/access_right/c_abf2Reconocimiento (by)http://creativecommons.org/licenses/by/4.0/info:eu-repo/semantics/openAccessoai:riunet.upv.es:10251/1932262026-06-13T07:49:27Z |
| dc.title.none.fl_str_mv |
Production of Potyvirus-Derived Nanoparticles Decorated with a Nanobody in Biofactory Plants |
| title |
Production of Potyvirus-Derived Nanoparticles Decorated with a Nanobody in Biofactory Plants |
| spellingShingle |
Production of Potyvirus-Derived Nanoparticles Decorated with a Nanobody in Biofactory Plants Martí, Mari Carmen Viral nanoparticle VNP Plant virus Nanobody Potyvirus Zucchini yellow mosaic virus Tobacco etch virus |
| title_short |
Production of Potyvirus-Derived Nanoparticles Decorated with a Nanobody in Biofactory Plants |
| title_full |
Production of Potyvirus-Derived Nanoparticles Decorated with a Nanobody in Biofactory Plants |
| title_fullStr |
Production of Potyvirus-Derived Nanoparticles Decorated with a Nanobody in Biofactory Plants |
| title_full_unstemmed |
Production of Potyvirus-Derived Nanoparticles Decorated with a Nanobody in Biofactory Plants |
| title_sort |
Production of Potyvirus-Derived Nanoparticles Decorated with a Nanobody in Biofactory Plants |
| dc.creator.none.fl_str_mv |
Martí, Mari Carmen Merwaiss, Fernando Butkovic, Anamarija Daròs, José-Antonio|||0000-0002-6535-2889 |
| author |
Martí, Mari Carmen |
| author_facet |
Martí, Mari Carmen Merwaiss, Fernando Butkovic, Anamarija Daròs, José-Antonio|||0000-0002-6535-2889 |
| author_role |
author |
| author2 |
Merwaiss, Fernando Butkovic, Anamarija Daròs, José-Antonio|||0000-0002-6535-2889 |
| author2_role |
author author author |
| dc.contributor.none.fl_str_mv |
Instituto Universitario Mixto de Biología Molecular y Celular de Plantas European Regional Development Fund Ministerio de Ciencia e Innovación Ministerio de Educación, Cultura y Deporte Repositorio Institucional de la Universitat Politècnica de València Riunet |
| dc.subject.none.fl_str_mv |
Viral nanoparticle VNP Plant virus Nanobody Potyvirus Zucchini yellow mosaic virus Tobacco etch virus |
| topic |
Viral nanoparticle VNP Plant virus Nanobody Potyvirus Zucchini yellow mosaic virus Tobacco etch virus |
| description |
[EN] Viral nanoparticles (VNPs) have recently attracted attention for their use as building blocks for novel materials to support a range of functions of potential interest in nanotechnology and medicine. Viral capsids are ideal for presenting small epitopes by inserting them at an appropriate site on the selected coat protein (CP). VNPs presenting antibodies on their surfaces are considered highly promising tools for therapeutic and diagnostic purposes. Due to their size, nanobodies are an interesting alternative to classic antibodies for surface presentation. Nanobodies are the variable domains of heavy-chain (VHH) antibodies from animals belonging to the family Camelidae, which have several properties that make them attractive therapeutic molecules, such as their small size, simple structure, and high affinity and specificity. In this work, we have produced genetically encoded VNPs derived from two different potyviruses¿the largest group of RNA viruses that infect plants¿decorated with nanobodies. We have created a VNP derived from zucchini yellow mosaic virus (ZYMV) decorated with a nanobody against the green fluorescent protein (GFP) in zucchini (Cucurbita pepo) plants. As reported for other viruses, the expression of ZYMV-derived VNPs decorated with this nanobody was only made possible by including a picornavirus 2A splicing peptide between the fused proteins, which resulted in a mixed population of unmodified and decorated CPs. We have also produced tobacco etch virus (TEV)-derived VNPs in Nicotiana benthamiana plants decorated with the same nanobody against GFP. Strikingly, in this case, VNPs could be assembled by direct fusion of the nanobody to the viral CP with no 2A splicing involved, likely resulting in fully decorated VNPs. For both expression systems, correct assembly and purification of the recombinant VNPs was confirmed by transmission electron microscope; the functionality of the CP-fused nanobody was assessed by western blot and binding assays. In sum, here we report the production of genetically encoded plant-derived VNPs decorated with a nanobody. This system may be an attractive alternative for the sustainable production in plants of nanobody-containing nanomaterials for diagnostic and therapeutic purposes. |
| publishDate |
2022 |
| dc.date.none.fl_str_mv |
2022 2022-03-31 |
| dc.type.none.fl_str_mv |
journal article http://purl.org/coar/resource_type/c_6501 VoR http://purl.org/coar/version/c_970fb48d4fbd8a85 |
| dc.type.openaire.fl_str_mv |
info:eu-repo/semantics/article |
| format |
article |
| dc.identifier.none.fl_str_mv |
https://riunet.upv.es/handle/10251/193226 |
| url |
https://riunet.upv.es/handle/10251/193226 |
| dc.language.none.fl_str_mv |
Inglés eng |
| language_invalid_str_mv |
Inglés |
| language |
eng |
| dc.relation.none.fl_str_mv |
Ministerio de Educación y Cultura http://dx.doi.org/10.13039/501100003176 FPU16%2F05294 FPU16%2F05294 Ministerio de Ciencia e Innovación http://dx.doi.org/10.13039/501100004837 PID2020-114691RB-I00 Biotecnología de virus de plantas: vectores virales y estrategias de resistencia |
| dc.rights.none.fl_str_mv |
open access http://purl.org/coar/access_right/c_abf2 Reconocimiento (by) http://creativecommons.org/licenses/by/4.0/ |
| dc.rights.openaire.fl_str_mv |
info:eu-repo/semantics/openAccess |
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open access http://purl.org/coar/access_right/c_abf2 Reconocimiento (by) http://creativecommons.org/licenses/by/4.0/ |
| eu_rights_str_mv |
openAccess |
| dc.format.none.fl_str_mv |
application/pdf |
| dc.publisher.none.fl_str_mv |
Frontiers Media S.A. |
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Frontiers Media S.A. |
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reponame:RiuNet. Repositorio Institucional de la Universitat Politécnica de Valéncia instname:Universitat Politècnica de València (UPV) |
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Universitat Politècnica de València (UPV) |
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RiuNet. Repositorio Institucional de la Universitat Politécnica de Valéncia |
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RiuNet. Repositorio Institucional de la Universitat Politécnica de Valéncia |
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