Gene-edited vero cells as rotavirus vaccine substrates.
CRISPR-Cas9
Enhanced vaccine cell line
Rotavirus
Vaccine substrates
Journal
Vaccine: X
ISSN: 2590-1362
Titre abrégé: Vaccine X
Pays: England
ID NLM: 101748769
Informations de publication
Date de publication:
10 Dec 2019
10 Dec 2019
Historique:
received:
04
06
2019
revised:
30
08
2019
accepted:
04
10
2019
entrez:
30
10
2019
pubmed:
30
10
2019
medline:
30
10
2019
Statut:
epublish
Résumé
Rotavirus (RV) is a leading cause of severe gastroenteritis globally and can cause substantial morbidity associated with gastroenteritis in children <5 years of age. Orally administered live-attenuated RV vaccines offer protection against disease but vaccination efforts have been hampered by high manufacturing costs and the need to maintain a cold chain. A subset of Vero cell host genes was identified by siRNA that when knocked down increased RV replication and these anti-viral host genes were individually deleted using CRISPR-Cas9. Fully-sequenced gene knockout Vero cell substrates were assessed for increased RV replication and RV vaccine antigen expression compared to wild type Vero cells. The results showed that RV replication and antigen production were logs higher in Vero cells having an We used siRNAs to screen for host genes that negatively affected RV replication, then CRISPR-Cas9 gene editing to delete select genes. The gene editing led to the development of enhanced RV vaccine substrates supporting a potential path forward for improving RV vaccine production.
Sections du résumé
BACKGROUND
BACKGROUND
Rotavirus (RV) is a leading cause of severe gastroenteritis globally and can cause substantial morbidity associated with gastroenteritis in children <5 years of age. Orally administered live-attenuated RV vaccines offer protection against disease but vaccination efforts have been hampered by high manufacturing costs and the need to maintain a cold chain.
METHODS
METHODS
A subset of Vero cell host genes was identified by siRNA that when knocked down increased RV replication and these anti-viral host genes were individually deleted using CRISPR-Cas9.
RESULTS
RESULTS
Fully-sequenced gene knockout Vero cell substrates were assessed for increased RV replication and RV vaccine antigen expression compared to wild type Vero cells. The results showed that RV replication and antigen production were logs higher in Vero cells having an
CONCLUSIONS
CONCLUSIONS
We used siRNAs to screen for host genes that negatively affected RV replication, then CRISPR-Cas9 gene editing to delete select genes. The gene editing led to the development of enhanced RV vaccine substrates supporting a potential path forward for improving RV vaccine production.
Identifiants
pubmed: 31660537
doi: 10.1016/j.jvacx.2019.100045
pii: S2590-1362(19)30046-4
pii: 100045
pmc: PMC6806661
doi:
Types de publication
Journal Article
Langues
eng
Pagination
100045Informations de copyright
© 2019 The Author(s).
Déclaration de conflit d'intérêts
The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: [The authors report no relationships or activities that could appear to have influenced the submitted work. CK has a patent on the RV3 rotavirus vaccine. The findings and conclusions in this report are those of the authors and do not necessarily represent the official positons of Centers for Disease Control and Prevention]
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