Baculovirus transduction in mammalian cells is affected by the production of type I and III interferons, which is mediated mainly by the cGAS-STING pathway

Autores
Amalfi, Sabrina; Molina, Guido Nicolas; Bevacqua, Romina Jimena; Lopez, Maria Gabriela; Taboga, Oscar Alberto; Alfonso, Victoria
Año de publicación
2020
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
The baculovirus Autographa californica multiple nucleopolyhedrovirus is an insect virus with a circular double-stranded DNA genome, which, among other multiple biotechnological applications, is used as an expression vector for gene delivery in mammalian cells. Nevertheless, the nonspecific immune response triggered by viral vectors often suppresses transgene expression. To understand the mechanisms involved in that response, in the present study, we studied the cyclic GMP-AMP synthase-stimulator of interferon genes (cGAS-STING) pathway by using two approaches: the genetic edition through CRISPR/Cas9 technology of genes encoding STING or cGAS in NIH/3T3 murine fibroblasts and the infection of HEK293 and HEK293 T human epithelial cells, deficient in cGAS and in cGAS and STING expression, respectively. Overall, our results suggest the existence of two different pathways involved in the establishment of the antiviral response, both dependent on STING expression. Particularly, the cGAS-STING pathway resulted in the more relevant production of beta interferon (IFN-β) and IFN-λ1 in response to baculovirus infection. In human epithelial cells, IFN-λ1 production was also induced in a cGAS-independent and DNA-protein kinase (DNA-PK)-dependent manner. Finally, we demonstrated that these cellular responses toward baculovirus infection affect the efficiency of transduction of baculovirus vectors. IMPORTANCE Baculoviruses are nonpathogenic viruses that infect mammals, which, among other applications, are used as vehicles for gene delivery. Here, we demonstrated that the cytosolic DNA sensor cGAS recognizes baculoviral DNA and that the cGAS-STING axis is primarily responsible for the attenuation of transduction in human and mouse cell lines through type I and type III IFNs. Furthermore, we identified DNA-dependent protein kinase (DNA-PK) as a cGAS-independent and alternative DNA cytosolic sensor that contributes less to the antiviral state in baculovirus infection in human epithelial cells than cGAS. Knowledge of the pathways involved in the response of mammalian cells to baculovirus infection will improve the use of this vector as a tool for gene therapy.
Instituto de Biotecnología
Fil: Amalfi, Sabrina. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Agrobiotecnología y Biología Molecular (IABIMO); Argentina
Fil: Amalfi, Sabrina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Molina, Guido Nicolas. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Agrobiotecnología y Biología Molecular (IABIMO); Argentina
Fil: Molina, Guido Nicolas. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Bevacqua, Romina Jimena. Universidad de Buenos Aires. Facultad de Agronomía. Laboratorio de Biotecnología Animal; Argentina
Fil: Bevacqua, Romina Jimena. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Bevacqua, Romina Jimena. Stanford University School of Medicine. Department of Developmental Biology. Seung Kim Lab; Estados Unidos
Fil: Lopez, Maria Gabriela. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Agrobiotecnología y Biología Molecular (IABIMO); Argentina
Fil: Lopez, Maria Gabriela. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Taboga, Oscar Alberto. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Agrobiotecnología y Biología Molecular (IABIMO); Argentina
Fil: Taboga, Oscar Alberto. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Alfonso, Victoria. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Agrobiotecnología y Biología Molecular (IABIMO); Argentina
Fil: Alfonso, Victoria. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fuente
Journal of Virology 94 (21) : e01555-20 (Noviembre 2020)
Materia
Baculovirus
Interferonas
Autographa californica
Repeticiones Palindrómicas Cortas Agrupadas y Regularmente Interespaciadas
Edición de Genes
Interferons
CRISPR
Gene Editing
Nivel de accesibilidad
acceso restringido
Condiciones de uso
Repositorio
INTA Digital (INTA)
Institución
Instituto Nacional de Tecnología Agropecuaria
OAI Identificador
oai:localhost:20.500.12123/10117
Acceder
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oai_identifier_str oai:localhost:20.500.12123/10117
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spelling Baculovirus transduction in mammalian cells is affected by the production of type I and III interferons, which is mediated mainly by the cGAS-STING pathwayAmalfi, SabrinaMolina, Guido NicolasBevacqua, Romina JimenaLopez, Maria GabrielaTaboga, Oscar AlbertoAlfonso, VictoriaBaculovirusInterferonasAutographa californicaRepeticiones Palindrómicas Cortas Agrupadas y Regularmente InterespaciadasEdición de GenesInterferonsCRISPRGene EditingThe baculovirus Autographa californica multiple nucleopolyhedrovirus is an insect virus with a circular double-stranded DNA genome, which, among other multiple biotechnological applications, is used as an expression vector for gene delivery in mammalian cells. Nevertheless, the nonspecific immune response triggered by viral vectors often suppresses transgene expression. To understand the mechanisms involved in that response, in the present study, we studied the cyclic GMP-AMP synthase-stimulator of interferon genes (cGAS-STING) pathway by using two approaches: the genetic edition through CRISPR/Cas9 technology of genes encoding STING or cGAS in NIH/3T3 murine fibroblasts and the infection of HEK293 and HEK293 T human epithelial cells, deficient in cGAS and in cGAS and STING expression, respectively. Overall, our results suggest the existence of two different pathways involved in the establishment of the antiviral response, both dependent on STING expression. Particularly, the cGAS-STING pathway resulted in the more relevant production of beta interferon (IFN-β) and IFN-λ1 in response to baculovirus infection. In human epithelial cells, IFN-λ1 production was also induced in a cGAS-independent and DNA-protein kinase (DNA-PK)-dependent manner. Finally, we demonstrated that these cellular responses toward baculovirus infection affect the efficiency of transduction of baculovirus vectors. IMPORTANCE Baculoviruses are nonpathogenic viruses that infect mammals, which, among other applications, are used as vehicles for gene delivery. Here, we demonstrated that the cytosolic DNA sensor cGAS recognizes baculoviral DNA and that the cGAS-STING axis is primarily responsible for the attenuation of transduction in human and mouse cell lines through type I and type III IFNs. Furthermore, we identified DNA-dependent protein kinase (DNA-PK) as a cGAS-independent and alternative DNA cytosolic sensor that contributes less to the antiviral state in baculovirus infection in human epithelial cells than cGAS. Knowledge of the pathways involved in the response of mammalian cells to baculovirus infection will improve the use of this vector as a tool for gene therapy.Instituto de BiotecnologíaFil: Amalfi, Sabrina. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Agrobiotecnología y Biología Molecular (IABIMO); ArgentinaFil: Amalfi, Sabrina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Molina, Guido Nicolas. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Agrobiotecnología y Biología Molecular (IABIMO); ArgentinaFil: Molina, Guido Nicolas. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Bevacqua, Romina Jimena. Universidad de Buenos Aires. Facultad de Agronomía. Laboratorio de Biotecnología Animal; ArgentinaFil: Bevacqua, Romina Jimena. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Bevacqua, Romina Jimena. Stanford University School of Medicine. Department of Developmental Biology. Seung Kim Lab; Estados UnidosFil: Lopez, Maria Gabriela. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Agrobiotecnología y Biología Molecular (IABIMO); ArgentinaFil: Lopez, Maria Gabriela. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Taboga, Oscar Alberto. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Agrobiotecnología y Biología Molecular (IABIMO); ArgentinaFil: Taboga, Oscar Alberto. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Alfonso, Victoria. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Agrobiotecnología y Biología Molecular (IABIMO); ArgentinaFil: Alfonso, Victoria. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaAmerican Society for Microbiology2021-08-25T17:58:49Z2021-08-25T17:58:49Z2020-11info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfhttp://hdl.handle.net/20.500.12123/10117https://journals.asm.org/doi/10.1128/JVI.01555-200022-538Xhttps://doi.org/10.1128/JVI.01555-20Journal of Virology 94 (21) : e01555-20 (Noviembre 2020)reponame:INTA Digital (INTA)instname:Instituto Nacional de Tecnología Agropecuariaenginfo:eu-repograntAgreement/INTA/PNBIO-1131034/AR./Inmunología molecular y genómica funcional aplicadas a interacciones patógeno hospedador de interés pecuario.info:eu-repo/semantics/restrictedAccess2025-09-29T13:45:19Zoai:localhost:20.500.12123/10117instacron:INTAInstitucionalhttp://repositorio.inta.gob.ar/Organismo científico-tecnológicoNo correspondehttp://repositorio.inta.gob.ar/oai/requesttripaldi.nicolas@inta.gob.arArgentinaNo correspondeNo correspondeNo correspondeopendoar:l2025-09-29 13:45:20.026INTA Digital (INTA) - Instituto Nacional de Tecnología Agropecuariafalse
dc.title.none.fl_str_mv Baculovirus transduction in mammalian cells is affected by the production of type I and III interferons, which is mediated mainly by the cGAS-STING pathway
title Baculovirus transduction in mammalian cells is affected by the production of type I and III interferons, which is mediated mainly by the cGAS-STING pathway
spellingShingle Baculovirus transduction in mammalian cells is affected by the production of type I and III interferons, which is mediated mainly by the cGAS-STING pathway
Amalfi, Sabrina
Baculovirus
Interferonas
Autographa californica
Repeticiones Palindrómicas Cortas Agrupadas y Regularmente Interespaciadas
Edición de Genes
Interferons
CRISPR
Gene Editing
title_short Baculovirus transduction in mammalian cells is affected by the production of type I and III interferons, which is mediated mainly by the cGAS-STING pathway
title_full Baculovirus transduction in mammalian cells is affected by the production of type I and III interferons, which is mediated mainly by the cGAS-STING pathway
title_fullStr Baculovirus transduction in mammalian cells is affected by the production of type I and III interferons, which is mediated mainly by the cGAS-STING pathway
title_full_unstemmed Baculovirus transduction in mammalian cells is affected by the production of type I and III interferons, which is mediated mainly by the cGAS-STING pathway
title_sort Baculovirus transduction in mammalian cells is affected by the production of type I and III interferons, which is mediated mainly by the cGAS-STING pathway
dc.creator.none.fl_str_mv Amalfi, Sabrina
Molina, Guido Nicolas
Bevacqua, Romina Jimena
Lopez, Maria Gabriela
Taboga, Oscar Alberto
Alfonso, Victoria
author Amalfi, Sabrina
author_facet Amalfi, Sabrina
Molina, Guido Nicolas
Bevacqua, Romina Jimena
Lopez, Maria Gabriela
Taboga, Oscar Alberto
Alfonso, Victoria
author_role author
author2 Molina, Guido Nicolas
Bevacqua, Romina Jimena
Lopez, Maria Gabriela
Taboga, Oscar Alberto
Alfonso, Victoria
author2_role author
author
author
author
author
dc.subject.none.fl_str_mv Baculovirus
Interferonas
Autographa californica
Repeticiones Palindrómicas Cortas Agrupadas y Regularmente Interespaciadas
Edición de Genes
Interferons
CRISPR
Gene Editing
topic Baculovirus
Interferonas
Autographa californica
Repeticiones Palindrómicas Cortas Agrupadas y Regularmente Interespaciadas
Edición de Genes
Interferons
CRISPR
Gene Editing
dc.description.none.fl_txt_mv The baculovirus Autographa californica multiple nucleopolyhedrovirus is an insect virus with a circular double-stranded DNA genome, which, among other multiple biotechnological applications, is used as an expression vector for gene delivery in mammalian cells. Nevertheless, the nonspecific immune response triggered by viral vectors often suppresses transgene expression. To understand the mechanisms involved in that response, in the present study, we studied the cyclic GMP-AMP synthase-stimulator of interferon genes (cGAS-STING) pathway by using two approaches: the genetic edition through CRISPR/Cas9 technology of genes encoding STING or cGAS in NIH/3T3 murine fibroblasts and the infection of HEK293 and HEK293 T human epithelial cells, deficient in cGAS and in cGAS and STING expression, respectively. Overall, our results suggest the existence of two different pathways involved in the establishment of the antiviral response, both dependent on STING expression. Particularly, the cGAS-STING pathway resulted in the more relevant production of beta interferon (IFN-β) and IFN-λ1 in response to baculovirus infection. In human epithelial cells, IFN-λ1 production was also induced in a cGAS-independent and DNA-protein kinase (DNA-PK)-dependent manner. Finally, we demonstrated that these cellular responses toward baculovirus infection affect the efficiency of transduction of baculovirus vectors. IMPORTANCE Baculoviruses are nonpathogenic viruses that infect mammals, which, among other applications, are used as vehicles for gene delivery. Here, we demonstrated that the cytosolic DNA sensor cGAS recognizes baculoviral DNA and that the cGAS-STING axis is primarily responsible for the attenuation of transduction in human and mouse cell lines through type I and type III IFNs. Furthermore, we identified DNA-dependent protein kinase (DNA-PK) as a cGAS-independent and alternative DNA cytosolic sensor that contributes less to the antiviral state in baculovirus infection in human epithelial cells than cGAS. Knowledge of the pathways involved in the response of mammalian cells to baculovirus infection will improve the use of this vector as a tool for gene therapy.
Instituto de Biotecnología
Fil: Amalfi, Sabrina. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Agrobiotecnología y Biología Molecular (IABIMO); Argentina
Fil: Amalfi, Sabrina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Molina, Guido Nicolas. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Agrobiotecnología y Biología Molecular (IABIMO); Argentina
Fil: Molina, Guido Nicolas. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Bevacqua, Romina Jimena. Universidad de Buenos Aires. Facultad de Agronomía. Laboratorio de Biotecnología Animal; Argentina
Fil: Bevacqua, Romina Jimena. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Bevacqua, Romina Jimena. Stanford University School of Medicine. Department of Developmental Biology. Seung Kim Lab; Estados Unidos
Fil: Lopez, Maria Gabriela. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Agrobiotecnología y Biología Molecular (IABIMO); Argentina
Fil: Lopez, Maria Gabriela. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Taboga, Oscar Alberto. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Agrobiotecnología y Biología Molecular (IABIMO); Argentina
Fil: Taboga, Oscar Alberto. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Alfonso, Victoria. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Agrobiotecnología y Biología Molecular (IABIMO); Argentina
Fil: Alfonso, Victoria. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
description The baculovirus Autographa californica multiple nucleopolyhedrovirus is an insect virus with a circular double-stranded DNA genome, which, among other multiple biotechnological applications, is used as an expression vector for gene delivery in mammalian cells. Nevertheless, the nonspecific immune response triggered by viral vectors often suppresses transgene expression. To understand the mechanisms involved in that response, in the present study, we studied the cyclic GMP-AMP synthase-stimulator of interferon genes (cGAS-STING) pathway by using two approaches: the genetic edition through CRISPR/Cas9 technology of genes encoding STING or cGAS in NIH/3T3 murine fibroblasts and the infection of HEK293 and HEK293 T human epithelial cells, deficient in cGAS and in cGAS and STING expression, respectively. Overall, our results suggest the existence of two different pathways involved in the establishment of the antiviral response, both dependent on STING expression. Particularly, the cGAS-STING pathway resulted in the more relevant production of beta interferon (IFN-β) and IFN-λ1 in response to baculovirus infection. In human epithelial cells, IFN-λ1 production was also induced in a cGAS-independent and DNA-protein kinase (DNA-PK)-dependent manner. Finally, we demonstrated that these cellular responses toward baculovirus infection affect the efficiency of transduction of baculovirus vectors. IMPORTANCE Baculoviruses are nonpathogenic viruses that infect mammals, which, among other applications, are used as vehicles for gene delivery. Here, we demonstrated that the cytosolic DNA sensor cGAS recognizes baculoviral DNA and that the cGAS-STING axis is primarily responsible for the attenuation of transduction in human and mouse cell lines through type I and type III IFNs. Furthermore, we identified DNA-dependent protein kinase (DNA-PK) as a cGAS-independent and alternative DNA cytosolic sensor that contributes less to the antiviral state in baculovirus infection in human epithelial cells than cGAS. Knowledge of the pathways involved in the response of mammalian cells to baculovirus infection will improve the use of this vector as a tool for gene therapy.
publishDate 2020
dc.date.none.fl_str_mv 2020-11
2021-08-25T17:58:49Z
2021-08-25T17:58:49Z
dc.type.none.fl_str_mv info:eu-repo/semantics/article
info:eu-repo/semantics/publishedVersion
http://purl.org/coar/resource_type/c_6501
info:ar-repo/semantics/articulo
format article
status_str publishedVersion
dc.identifier.none.fl_str_mv http://hdl.handle.net/20.500.12123/10117
https://journals.asm.org/doi/10.1128/JVI.01555-20
0022-538X
https://doi.org/10.1128/JVI.01555-20
url http://hdl.handle.net/20.500.12123/10117
https://journals.asm.org/doi/10.1128/JVI.01555-20
https://doi.org/10.1128/JVI.01555-20
identifier_str_mv 0022-538X
dc.language.none.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv info:eu-repograntAgreement/INTA/PNBIO-1131034/AR./Inmunología molecular y genómica funcional aplicadas a interacciones patógeno hospedador de interés pecuario.
dc.rights.none.fl_str_mv info:eu-repo/semantics/restrictedAccess
eu_rights_str_mv restrictedAccess
dc.format.none.fl_str_mv application/pdf
dc.publisher.none.fl_str_mv American Society for Microbiology
publisher.none.fl_str_mv American Society for Microbiology
dc.source.none.fl_str_mv Journal of Virology 94 (21) : e01555-20 (Noviembre 2020)
reponame:INTA Digital (INTA)
instname:Instituto Nacional de Tecnología Agropecuaria
reponame_str INTA Digital (INTA)
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instname_str Instituto Nacional de Tecnología Agropecuaria
repository.name.fl_str_mv INTA Digital (INTA) - Instituto Nacional de Tecnología Agropecuaria
repository.mail.fl_str_mv tripaldi.nicolas@inta.gob.ar
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