Robust Enhancement of Lentivirus Production by Promoter Activation

Abstract Lentiviral vectors are a valuable tool to deliver exogenous genes for stable expression in cells. While much progress has been made in processing lentiviral vector-containing culture medium, it remains to be explored how the production of lentiviral vector from producer cells can be increas...

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Autores principales:	Naoto Suzuki, Takeshi Yoshida, Hiroaki Takeuchi, Ryuta Sakuma, Sayaka Sukegawa, Shoji Yamaoka
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2018
Materias:	Lentiviral Vector Production Really Interesting New Gene (RING) SPRY Domain Expressing Firefly Luciferase Vesicular Stomatitis Virus Glycoprotein (VSV-G) Medicine R Science Q
Acceso en línea:	https://doaj.org/article/e880902b6ac14fb2b80677fa0462439c
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spelling	oai:doaj.org-article:e880902b6ac14fb2b80677fa0462439c2021-12-02T15:08:18ZRobust Enhancement of Lentivirus Production by Promoter Activation10.1038/s41598-018-33042-52045-2322https://doaj.org/article/e880902b6ac14fb2b80677fa0462439c2018-10-01T00:00:00Zhttps://doi.org/10.1038/s41598-018-33042-5https://doaj.org/toc/2045-2322Abstract Lentiviral vectors are a valuable tool to deliver exogenous genes for stable expression in cells. While much progress has been made in processing lentiviral vector-containing culture medium, it remains to be explored how the production of lentiviral vector from producer cells can be increased. We initially found that co-expression of the SPRY domain-containing SOCS box protein 1 (SPSB1) promotes the production of human immunodeficiency virus type 1 (HIV-1) and lentiviral vector with increased expression of the Gag and envelope proteins and activation of the HIV-1 LTR and CMV promoter. The presence of AP-1, NF-κB and CREB/ATF recognition sites in these promoters prompted us to utilize human T-lymphotropic virus type 1 (HTLV-1) Tax for lentiviral vector production because Tax activates all these transcription factors. Co-expression of a small amount of Tax markedly increased both the expression of viral structural proteins in producer cells and release of lentiviral vector particles, resulting in a more than 10-fold enhancement of transduction efficiency. Of note, the Tax protein was not detected in the lentiviral vector particles concentrated by ultracentrifugation, supporting the safety of this preparation. Collectively, these results indicate that promoter activation in producer cells represents a promising approach to preparing high-titer lentiviral vectors.Naoto SuzukiTakeshi YoshidaHiroaki TakeuchiRyuta SakumaSayaka SukegawaShoji YamaokaNature PortfolioarticleLentiviral Vector ProductionReally Interesting New Gene (RING)SPRY DomainExpressing Firefly LuciferaseVesicular Stomatitis Virus Glycoprotein (VSV-G)MedicineRScienceQENScientific Reports, Vol 8, Iss 1, Pp 1-9 (2018)
institution	DOAJ
collection	DOAJ
language	EN
topic	Lentiviral Vector Production Really Interesting New Gene (RING) SPRY Domain Expressing Firefly Luciferase Vesicular Stomatitis Virus Glycoprotein (VSV-G) Medicine R Science Q
spellingShingle	Lentiviral Vector Production Really Interesting New Gene (RING) SPRY Domain Expressing Firefly Luciferase Vesicular Stomatitis Virus Glycoprotein (VSV-G) Medicine R Science Q Naoto Suzuki Takeshi Yoshida Hiroaki Takeuchi Ryuta Sakuma Sayaka Sukegawa Shoji Yamaoka Robust Enhancement of Lentivirus Production by Promoter Activation
description	Abstract Lentiviral vectors are a valuable tool to deliver exogenous genes for stable expression in cells. While much progress has been made in processing lentiviral vector-containing culture medium, it remains to be explored how the production of lentiviral vector from producer cells can be increased. We initially found that co-expression of the SPRY domain-containing SOCS box protein 1 (SPSB1) promotes the production of human immunodeficiency virus type 1 (HIV-1) and lentiviral vector with increased expression of the Gag and envelope proteins and activation of the HIV-1 LTR and CMV promoter. The presence of AP-1, NF-κB and CREB/ATF recognition sites in these promoters prompted us to utilize human T-lymphotropic virus type 1 (HTLV-1) Tax for lentiviral vector production because Tax activates all these transcription factors. Co-expression of a small amount of Tax markedly increased both the expression of viral structural proteins in producer cells and release of lentiviral vector particles, resulting in a more than 10-fold enhancement of transduction efficiency. Of note, the Tax protein was not detected in the lentiviral vector particles concentrated by ultracentrifugation, supporting the safety of this preparation. Collectively, these results indicate that promoter activation in producer cells represents a promising approach to preparing high-titer lentiviral vectors.
format	article
author	Naoto Suzuki Takeshi Yoshida Hiroaki Takeuchi Ryuta Sakuma Sayaka Sukegawa Shoji Yamaoka
author_facet	Naoto Suzuki Takeshi Yoshida Hiroaki Takeuchi Ryuta Sakuma Sayaka Sukegawa Shoji Yamaoka
author_sort	Naoto Suzuki
title	Robust Enhancement of Lentivirus Production by Promoter Activation
title_short	Robust Enhancement of Lentivirus Production by Promoter Activation
title_full	Robust Enhancement of Lentivirus Production by Promoter Activation
title_fullStr	Robust Enhancement of Lentivirus Production by Promoter Activation
title_full_unstemmed	Robust Enhancement of Lentivirus Production by Promoter Activation
title_sort	robust enhancement of lentivirus production by promoter activation
publisher	Nature Portfolio
publishDate	2018
url	https://doaj.org/article/e880902b6ac14fb2b80677fa0462439c
work_keys_str_mv	AT naotosuzuki robustenhancementoflentivirusproductionbypromoteractivation AT takeshiyoshida robustenhancementoflentivirusproductionbypromoteractivation AT hiroakitakeuchi robustenhancementoflentivirusproductionbypromoteractivation AT ryutasakuma robustenhancementoflentivirusproductionbypromoteractivation AT sayakasukegawa robustenhancementoflentivirusproductionbypromoteractivation AT shojiyamaoka robustenhancementoflentivirusproductionbypromoteractivation
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Robust Enhancement of Lentivirus Production by Promoter Activation

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