Factors affecting rAAV titers during triple-plasmid transient transfection in HEK-293 cells.
Gene Therapy
HEK-293 cells
Plasmid tracking
Transient transfection
Virus production
rAAV
Journal
Biotechnology letters
ISSN: 1573-6776
Titre abrégé: Biotechnol Lett
Pays: Netherlands
ID NLM: 8008051
Informations de publication
Date de publication:
11 Sep 2024
11 Sep 2024
Historique:
received:
18
01
2024
accepted:
06
08
2024
revised:
24
06
2024
medline:
11
9
2024
pubmed:
11
9
2024
entrez:
11
9
2024
Statut:
aheadofprint
Résumé
The efficiency of triple-plasmid transfection in recombinant Adeno-Associated Virus (rAAV) production was analyzed by examining two distinct HEK-293 cells lines. These were categorized as high producer (HP) and low producer (LP) based on their differing levels of productivity under identical conditions. Analysis of RNA expression levels of viral genes revealed disparities in plasmid derived gene expression between the cell lines. Further assessment of transfection efficiency utilizing labeled plasmids revealed lower plasmid uptake and less efficient nuclear transport in LP cell line. Additionally, we observed inferior translation activity in LP, contributing to its shortcomings in overall productivity. In our attempt to optimize plasmid ratios to enhance fully packaged rAAV particle yield, we discovered cell-line-specific optimization potential. The findings highlight the transfection's complexity, urging tailored strategies for improved rAAV production based on each cell line's characteristics, enhancing understanding and guiding further efficiency optimization in rAAV production.
Identifiants
pubmed: 39259435
doi: 10.1007/s10529-024-03520-0
pii: 10.1007/s10529-024-03520-0
doi:
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Informations de copyright
© 2024. The Author(s).
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