清脆的
生物
遗传学
轮状病毒
钥匙(锁)
病毒学
寄主(生物学)
反向遗传学
生物技术
基因
病毒
基因组
生态学
作者
Yinxing Zhu,Meagan E. Sullender,Danielle E. Campbell,Leran Wang,Sanghyun Lee,Takahiro Kawagishi,Gaopeng Hou,Alen Dizdarevic,Philippe Jaïs,Megan T. Baldridge,Siyuan Ding
出处
期刊:npj vaccines
[Nature Portfolio]
日期:2024-11-06
卷期号:9 (1)
被引量:1
标识
DOI:10.1038/s41541-024-01007-7
摘要
Rotaviruses pose a significant threat to young children. To identify novel pro- and anti-rotavirus host factors, we performed genome-wide CRISPR/Cas9 screens using rhesus rotavirus and African green monkey cells. Genetic deletion of either SERPINB1 or TMEM236, the top two antiviral factors, in MA104 cells increased virus titers in a rotavirus strain independent manner. Using this information, we optimized the existing rotavirus reverse genetics systems by combining SERPINB1 knockout MA104 cells with a C3P3-G3 helper plasmid. We improved the recovery efficiency and rescued several low-titer rotavirus reporter and mutant strains that prove difficult to rescue otherwise. Furthermore, we demonstrate that TMEM236 knockout in Vero cells supported higher yields of two live-attenuated rotavirus vaccine strains than the parental cell line and represents a more robust vaccine-producing cell substrate. Collectively, we developed a third-generation optimized rotavirus reverse genetics system and generated gene-edited Vero cells as a new substrate for improving rotavirus vaccine production.
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