CRISPR screen to identify genes that contribute to type I interferon restriction of Zika virus

We performed CRISPR screens to determine human genes that contribute to type I interferon restriction of Zika virus in A549 cells. We used the PIKA CRISPR library (Ohainle et al., eLife 2018). All conditions included a 24-hour pre-treatment with 1000U/mL of IFN-beta. Cells were then infected with ZIKV (PRVABC59) at MOI=3 or left uninfected as the control condition. Dead/dying cells were harvested at 42 hours post infection by collecting the cell supernatant. Two technical replicates were performed for both the uninfected and infected conditions, and we performed the screen twice, resulting in 8 total samples.

本研究通过CRISPR筛选（CRISPR screen），以鉴定A549细胞中参与I型干扰素（type I interferon）限制寨卡病毒（Zika virus）增殖的人类基因。本次筛选使用PIKA CRISPR文库（Ohainle等人，《eLife》，2018年）。所有实验组均采用1000U/mL的β干扰素（IFN-β）进行24小时预处理。随后将细胞以感染复数（multiplicity of infection, MOI）=3的比例感染寨卡病毒（ZIKV, PRVABC59株），同时设置未感染组作为对照。于感染后42小时收集细胞上清液，以获取死亡/濒死细胞。未感染组与感染组均设置2次技术重复，且本筛选共开展2轮独立实验，最终共计获得8份样本。