Characterization and mutagenesis of Chinese hamster ovary cells endogenous retroviruses to inactivate viral particle release.

Details

Serval ID
serval:BIB_3F6D075DCCE1
Type
Article: article from journal or magazin.
Collection
Publications
Institution
Title
Characterization and mutagenesis of Chinese hamster ovary cells endogenous retroviruses to inactivate viral particle release.
Journal
Biotechnology and bioengineering
Author(s)
Duroy P.O., Bosshard S., Schmid-Siegert E., Neuenschwander S., Arib G., Lemercier P., Masternak J., Roesch L., Buron F., Girod P.A., Xenarios I., Mermod N.
ISSN
1097-0290 (Electronic)
ISSN-L
0006-3592
Publication state
Published
Issued date
02/2020
Peer-reviewed
Oui
Volume
117
Number
2
Pages
466-485
Language
english
Notes
Publication types: Journal Article
Publication Status: ppublish
Abstract
The Chinese hamster ovary (CHO) cells used to produce biopharmaceutical proteins are known to contain type-C endogenous retrovirus (ERV) sequences in their genome and to release retroviral-like particles. Although evidence for their infectivity is missing, this has raised safety concerns. As the genomic origin of these particles remained unclear, we characterized type-C ERV elements at the genome, transcriptome, and viral particle RNA levels. We identified 173 type-C ERV sequences clustering into three functionally conserved groups. Transcripts from one type-C ERV group were full-length, with intact open reading frames, and cognate viral genome RNA was loaded into retroviral-like particles, suggesting that this ERV group may produce functional viruses. CRISPR-Cas9 genome editing was used to disrupt the gag gene of the expressed type-C ERV group. Comparison of CRISPR-derived mutations at the DNA and RNA level led to the identification of a single ERV as the main source of the release of RNA-loaded viral particles. Clones bearing a Gag loss-of-function mutation in this ERV showed a reduction of RNA-containing viral particle release down to detection limits, without compromising cell growth or therapeutic protein production. Overall, our study provides a strategy to mitigate potential viral particle contaminations resulting from ERVs during biopharmaceutical manufacturing.
Keywords
Adventitious agents, CHO cells, Endogenous retroviral elements, Genome editing, Chinese hamster ovary cells, adventitious agents, endogenous retroviral elements, genome editing
Pubmed
Web of science
Open Access
Yes
Create date
24/10/2019 17:57
Last modification date
05/02/2020 7:19
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