Facile method for delivering chikungunya viral replicons into mosquitoes and mammalian cells

Abstract Reverse genetics is an important tool in the elucidation of viral replication and the development of countermeasures; however, these methods are impeded by laborious and inefficient replicon delivery methods. This paper demonstrates the use of a baculovirus to facilitate the efficient deliv...

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Autores principales:	Hui-Chung Lin, Der-Jiang Chiao, Chang-Chi Lin, Szu-Cheng Kuo
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2021
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/168163da6cbb41e387aadf6f6b024594
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spelling	oai:doaj.org-article:168163da6cbb41e387aadf6f6b0245942021-12-02T17:52:22ZFacile method for delivering chikungunya viral replicons into mosquitoes and mammalian cells10.1038/s41598-021-91830-y2045-2322https://doaj.org/article/168163da6cbb41e387aadf6f6b0245942021-06-01T00:00:00Zhttps://doi.org/10.1038/s41598-021-91830-yhttps://doaj.org/toc/2045-2322Abstract Reverse genetics is an important tool in the elucidation of viral replication and the development of countermeasures; however, these methods are impeded by laborious and inefficient replicon delivery methods. This paper demonstrates the use of a baculovirus to facilitate the efficient delivery of autonomous CHIKV replicons into mosquito and mammalian cells in vitro as well as adult mosquitoes in vivo. The efficacy of this approach was verified via co-localization among an eGFP reporter, nsP1, and dsRNA as well as through the inhibition of an RNA-dependent RNA polymerase (RdRp) null mutation (DDAA) in nsP4, or the treatment of a known antiviral compound (6-azauridine). We also investigated the correlation between CHIKV replicon-launched eGFP expression and the effectiveness of CHIKV replicon variants in inducing IFN-β expression in human cell lines. This delivery method based on a single vector is applicable to mosquito and mammalian cells in seeking to decipher the mechanisms underlying CHIKV replication, elucidate virus–host interactions, and develop antivirals. This study presents an effective alternative to overcome many of the technological issues related to the study and utilization of autonomous arbovirus replicons.Hui-Chung LinDer-Jiang ChiaoChang-Chi LinSzu-Cheng KuoNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 11, Iss 1, Pp 1-12 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Hui-Chung Lin Der-Jiang Chiao Chang-Chi Lin Szu-Cheng Kuo Facile method for delivering chikungunya viral replicons into mosquitoes and mammalian cells
description	Abstract Reverse genetics is an important tool in the elucidation of viral replication and the development of countermeasures; however, these methods are impeded by laborious and inefficient replicon delivery methods. This paper demonstrates the use of a baculovirus to facilitate the efficient delivery of autonomous CHIKV replicons into mosquito and mammalian cells in vitro as well as adult mosquitoes in vivo. The efficacy of this approach was verified via co-localization among an eGFP reporter, nsP1, and dsRNA as well as through the inhibition of an RNA-dependent RNA polymerase (RdRp) null mutation (DDAA) in nsP4, or the treatment of a known antiviral compound (6-azauridine). We also investigated the correlation between CHIKV replicon-launched eGFP expression and the effectiveness of CHIKV replicon variants in inducing IFN-β expression in human cell lines. This delivery method based on a single vector is applicable to mosquito and mammalian cells in seeking to decipher the mechanisms underlying CHIKV replication, elucidate virus–host interactions, and develop antivirals. This study presents an effective alternative to overcome many of the technological issues related to the study and utilization of autonomous arbovirus replicons.
format	article
author	Hui-Chung Lin Der-Jiang Chiao Chang-Chi Lin Szu-Cheng Kuo
author_facet	Hui-Chung Lin Der-Jiang Chiao Chang-Chi Lin Szu-Cheng Kuo
author_sort	Hui-Chung Lin
title	Facile method for delivering chikungunya viral replicons into mosquitoes and mammalian cells
title_short	Facile method for delivering chikungunya viral replicons into mosquitoes and mammalian cells
title_full	Facile method for delivering chikungunya viral replicons into mosquitoes and mammalian cells
title_fullStr	Facile method for delivering chikungunya viral replicons into mosquitoes and mammalian cells
title_full_unstemmed	Facile method for delivering chikungunya viral replicons into mosquitoes and mammalian cells
title_sort	facile method for delivering chikungunya viral replicons into mosquitoes and mammalian cells
publisher	Nature Portfolio
publishDate	2021
url	https://doaj.org/article/168163da6cbb41e387aadf6f6b024594
work_keys_str_mv	AT huichunglin facilemethodfordeliveringchikungunyaviralrepliconsintomosquitoesandmammaliancells AT derjiangchiao facilemethodfordeliveringchikungunyaviralrepliconsintomosquitoesandmammaliancells AT changchilin facilemethodfordeliveringchikungunyaviralrepliconsintomosquitoesandmammaliancells AT szuchengkuo facilemethodfordeliveringchikungunyaviralrepliconsintomosquitoesandmammaliancells
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Facile method for delivering chikungunya viral replicons into mosquitoes and mammalian cells

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