Gene Expression Analysis of Adapted Insect Cells during Influenza VLP Production Using RNA-Sequencing
Adaptive laboratory evolution has been used to improve production of influenza hemagglutinin (HA)-displaying virus-like particles (VLPs) in insect cells. However, little is known about the underlying biological mechanisms promoting higher HA-VLP expression in such adapted cell lines. In this article...
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MDPI AG
2022-10-01
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| Online Access: | https://www.mdpi.com/1999-4915/14/10/2238 |
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| author | Marco Silvano Ricardo Correia Nikolaus Virgolini Colin Clarke Paula M. Alves Inês A. Isidro António Roldão |
| author_facet | Marco Silvano Ricardo Correia Nikolaus Virgolini Colin Clarke Paula M. Alves Inês A. Isidro António Roldão |
| author_sort | Marco Silvano |
| collection | DOAJ |
| description | Adaptive laboratory evolution has been used to improve production of influenza hemagglutinin (HA)-displaying virus-like particles (VLPs) in insect cells. However, little is known about the underlying biological mechanisms promoting higher HA-VLP expression in such adapted cell lines. In this article, we present a study of gene expression patterns associated with high-producer insect High Five cells adapted to neutral pH, in comparison to non-adapted cells, during expression of influenza HA-VLPs. RNA-seq shows a decrease in the amount of reads mapping to host cell genomes along infection, and an increase in those mapping to baculovirus and transgenes. A total of 1742 host cell genes were found differentially expressed between adapted and non-adapted cells throughout infection, 474 of those being either up- or down-regulated at both time points evaluated (12 and 24 h post-infection). Interestingly, while host cell genes were found up- and down-regulated in an approximately 1:1 ratio, all differentially expressed baculovirus genes were found to be down-regulated in infected adapted cells. Pathway analysis of differentially expressed genes revealed enrichment of ribosome biosynthesis and carbohydrate, amino acid, and lipid metabolism. In addition, oxidative phosphorylation and protein folding, sorting and degradation pathways were also found to be overrepresented. These findings contribute to our knowledge of biological mechanisms of insect cells during baculovirus-mediated transient expression and will assist the identification of potential engineering targets to increase recombinant protein production in the future. |
| first_indexed | 2024-03-09T19:22:42Z |
| format | Article |
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| institution | Directory Open Access Journal |
| issn | 1999-4915 |
| language | English |
| last_indexed | 2024-03-09T19:22:42Z |
| publishDate | 2022-10-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Viruses |
| spelling | doaj.art-2afe4b70b00b4b66b68060250bf9ce472023-11-24T03:09:57ZengMDPI AGViruses1999-49152022-10-011410223810.3390/v14102238Gene Expression Analysis of Adapted Insect Cells during Influenza VLP Production Using RNA-SequencingMarco Silvano0Ricardo Correia1Nikolaus Virgolini2Colin Clarke3Paula M. Alves4Inês A. Isidro5António Roldão6IBET, Instituto de Biologia Experimental e Tecnológica, Apartado 12, 2780-901 Oeiras, PortugalIBET, Instituto de Biologia Experimental e Tecnológica, Apartado 12, 2780-901 Oeiras, PortugalIBET, Instituto de Biologia Experimental e Tecnológica, Apartado 12, 2780-901 Oeiras, PortugalNIBRT, National Institute for Bioprocessing Research and Training, Fosters Avenue, Blackrock, Co., A94 X099 Dublin, IrelandIBET, Instituto de Biologia Experimental e Tecnológica, Apartado 12, 2780-901 Oeiras, PortugalIBET, Instituto de Biologia Experimental e Tecnológica, Apartado 12, 2780-901 Oeiras, PortugalIBET, Instituto de Biologia Experimental e Tecnológica, Apartado 12, 2780-901 Oeiras, PortugalAdaptive laboratory evolution has been used to improve production of influenza hemagglutinin (HA)-displaying virus-like particles (VLPs) in insect cells. However, little is known about the underlying biological mechanisms promoting higher HA-VLP expression in such adapted cell lines. In this article, we present a study of gene expression patterns associated with high-producer insect High Five cells adapted to neutral pH, in comparison to non-adapted cells, during expression of influenza HA-VLPs. RNA-seq shows a decrease in the amount of reads mapping to host cell genomes along infection, and an increase in those mapping to baculovirus and transgenes. A total of 1742 host cell genes were found differentially expressed between adapted and non-adapted cells throughout infection, 474 of those being either up- or down-regulated at both time points evaluated (12 and 24 h post-infection). Interestingly, while host cell genes were found up- and down-regulated in an approximately 1:1 ratio, all differentially expressed baculovirus genes were found to be down-regulated in infected adapted cells. Pathway analysis of differentially expressed genes revealed enrichment of ribosome biosynthesis and carbohydrate, amino acid, and lipid metabolism. In addition, oxidative phosphorylation and protein folding, sorting and degradation pathways were also found to be overrepresented. These findings contribute to our knowledge of biological mechanisms of insect cells during baculovirus-mediated transient expression and will assist the identification of potential engineering targets to increase recombinant protein production in the future.https://www.mdpi.com/1999-4915/14/10/2238insect cellsbaculovirus expression systeminfluenza VLPRNA sequencingpathway analysis |
| spellingShingle | Marco Silvano Ricardo Correia Nikolaus Virgolini Colin Clarke Paula M. Alves Inês A. Isidro António Roldão Gene Expression Analysis of Adapted Insect Cells during Influenza VLP Production Using RNA-Sequencing Viruses insect cells baculovirus expression system influenza VLP RNA sequencing pathway analysis |
| title | Gene Expression Analysis of Adapted Insect Cells during Influenza VLP Production Using RNA-Sequencing |
| title_full | Gene Expression Analysis of Adapted Insect Cells during Influenza VLP Production Using RNA-Sequencing |
| title_fullStr | Gene Expression Analysis of Adapted Insect Cells during Influenza VLP Production Using RNA-Sequencing |
| title_full_unstemmed | Gene Expression Analysis of Adapted Insect Cells during Influenza VLP Production Using RNA-Sequencing |
| title_short | Gene Expression Analysis of Adapted Insect Cells during Influenza VLP Production Using RNA-Sequencing |
| title_sort | gene expression analysis of adapted insect cells during influenza vlp production using rna sequencing |
| topic | insect cells baculovirus expression system influenza VLP RNA sequencing pathway analysis |
| url | https://www.mdpi.com/1999-4915/14/10/2238 |
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