@article{6351628210ec40d4b7501f55f5f51a08,
title = "Gene-edited vero cells as rotavirus vaccine substrates",
abstract = "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: 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: 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 EMX2 gene deletion compared to other Vero cell substrates tested. 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.",
keywords = "CRISPR-Cas9, Enhanced vaccine cell line, Rotavirus, Vaccine substrates",
author = "Nichole Orr-Burks and Jackelyn Murray and Weilin Wu and Kirkwood, {Carl D.} and Todd, {Kyle V.} and Les Jones and Abhijeet Bakre and Houping Wang and Baoming Jiang and Tripp, {Ralph A.}",
note = "Funding Information: We would like to thank the Georgia Research Alliance, MCRI Australia, the members of the CDC Viral Gastroenteritis Branch for reagents, assays, and interpreting results and assistance from Arjun N. Bhatt and the University of Georgia Undergraduate Research Opportunities (CURO). We declare there are no conflicts of interest. All authors were involved in the design of the studies. All authors contributed to drafts and approved the final manuscript. The views expressed are those of the author(s). The funders had no role in the study design, data collection, analysis, or interpretation. This research was funded in part by the Bill and Melinda Gates Foundation, OPP1097219 to RT, the Georgia Research Alliance to RT, and the CDC/UGA Collaboration seed grant with BJ and RT. Funding Information: This research was funded in part by the Bill and Melinda Gates Foundation , OPP1097219 to RT, the Georgia Research Alliance to RT , and the CDC/UGA Collaboration seed grant with BJ and RT. Appendix A Publisher Copyright: {\textcopyright} 2019 The Author(s)",
year = "2019",
month = dec,
day = "10",
doi = "10.1016/j.jvacx.2019.100045",
language = "English",
volume = "3",
pages = "1--11",
journal = "Vaccine: X",
issn = "2590-1362",
publisher = "Elsevier",
}
