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使用CRISPR/Cas9构建IFNAR1淘汰MDBK细胞系及其对牛病毒复制的影响

Yuanchen Geng^1,2,3, Chuanwen Jiang^1,2,3, Hao Yang^1,2,3

¹State Key Laboratory of Agricultural Microbiology, Hubei Hongshan Laboratory, College of Veterinary Medicine, Huazhong Agricultural University, Wuhan, China.

Frontiers in immunology

|August 5, 2024

概括

研究人员创建了一个缺少IFNAR1基因的淘汰细胞系,以增强牛病毒复制. 这种IFNAR1淘汰MDBK细胞系支持增加病毒标位,为研究牛病毒和免疫反应提供了有价值的工具.

科学领域:

病毒学病毒学
免疫学免疫学免疫学
分子生物学分子生物学
遗传学遗传学是一个

背景情况:

I型干扰素 (IFN) 途径对抗病毒感染的细胞防御至关重要,但可以限制病毒复制.
了解宿主-病原体相互作用需要强大的模型系统来研究病毒复制和先天免疫力.

研究的目的:

使用CRISPR/Cas9技术生成IFNAR1淘汰 (KO) 的马丁-达比牛 (MDBK) 细胞系.
调查IFNAR1 KO MDBK细胞系对增强牛病毒复制的有用性,并阐明其潜在机制.

主要方法:

使用CRISPR/Cas9基因编辑来创建IFNAR1淘汰MDBK细胞系.
评估了牛病毒性腹病毒 (BVDV),牛肠道病毒和3型牛甲型流感病毒的病毒复制量.
进行RNA测序 (RNA-seq) 来分析野生型 (WT) 和IFNAR1KO细胞中的基因表达变化.

主要成果:

IFNAR1 KO MDBK细胞对BVDV (1.5 log10) 和其他牛病毒的标位显著增加.
RNA-seq分析揭示了IFN-I通路基因和与KO细胞中信号传递和细胞周期调节相关的差异表达基因 (DEGs) 的改变表达.
与单个IFNAR1 KO.相比,双淘汰细胞系 (IFNAR1/IFNAR2) 进一步增强了BVDV复制.

结论:

开发的IFNAR1 KO MDBK细胞系是改善各种牛病毒复制的有效模型.
这一细胞系为推进对牛病毒病原和宿主天生的免疫反应的研究提供了宝贵的工具.

关键词:

在 BVDVV 的位置上.这就是CRISPR/Cas9的作用.在IFNAR1中,这就是IL13RA2的原因.在MDBK细胞系中,在 SLC25A3434 中使用.

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