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通过CRISPR/Cas9选,可以识别增强轮回病毒逆转基因功效和疫苗生产的关键宿主因素

Yinxing Zhu¹, Meagan E Sullender², Danielle E Campbell²

¹Department of Molecular Microbiology, Washington University School of Medicine, St. Louis, MO, USA.

|November 6, 2024

概括

研究人员将SERPINB1和TMEM236确定为对抗轮状病毒的关键抗病毒因素. 通过这些基因淘汰来增强细胞系,可以改善轮状病毒的恢复和疫苗生产效率.

科学领域:

病毒学病毒学
遗传学遗传学是一个

背景情况:

罗塔病毒是全球婴儿严重腹的主要原因.
识别影响轮状病毒复制的宿主因素对于治疗和疫苗开发至关重要.

研究的目的:

通过全基因组CRISPR屏幕发现影响轮状病毒复制的新型宿主因素.
为了优化轮状病毒逆转基因系统,并开发改善的细胞基质用于疫苗生产.

主要方法:

在带有 rhesus 轮状病毒的非洲绿细胞 (MA104) 中进行全基因组的 CRISPR/Cas9 查.
在MA104和Vero细胞中发现的抗病毒因子 (SERPINB1,TMEM236) 的遗传删除.
使用SERPINB1淘汰细胞优化轮状病毒逆遗传系统.
评估TMEM236淘汰赛Vero细胞用于生衰减轮状病毒疫苗生产.

主要成果:

鉴定出SERPINB1和TMEM236是强大的抗病毒因子,它们的删除会增加轮状病毒标位.
开发了一种优化的第三代轮状病毒逆遗传系统,增强了难以恢复的菌株.
与父细胞相比,TMEM236淘汰赛Vero细胞对活衰减的轮状病毒疫苗菌株的产量明显更高.

结论:

在控制轮状病毒感染方面,SERPINB1和TMEM236是关键的宿主因素.
开发的逆转基因系统和基因编辑的Vero细胞代表了轮状病毒研究和疫苗制造的重大进步.

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