CRISPR screen for interferon-alpha inducible inhibitors of yellow fever virus replication

Host cells produce interferon (IFN) in response to viral infections. Secreted interferon results in the transcription and production of hundreds of interferon-stimulated genes (ISGs). A genome-wide CRISPR screen using IFN alpha-treated Huh7.5 cells was performed to determine which ISGs were required in order for host cells to suppress yellow fever virus (YFV) infection. Overall design: Human Huh7.5 cells were transduced with a human CRISPR library in a single replicate at 900X library coverage, treated with IFN alpha, infected with YFV-17D-venus, and GFP-positive cells that escaped IFN-alpha suppression of YFV-17D-venus infection were collected by FACS.宿主细胞在遭遇病毒感染时会产生干扰素（interferon, IFN）。分泌至胞外的干扰素可诱导数百个干扰素刺激基因（interferon-stimulated genes, ISGs）的转录与表达。本研究以经α干扰素处理的Huh7.5细胞为模型开展全基因组CRISPR筛选，旨在明确宿主细胞抑制黄热病毒（yellow fever virus, YFV）感染所必需的干扰素刺激基因。实验总体设计如下：将人源CRISPR文库以900倍文库覆盖度进行单重复转导，随后用α干扰素处理细胞并感染YFV-17D-venus毒株，最终通过荧光激活细胞分选（FACS）收集逃脱α干扰素介导的YFV-17D-venus感染抑制的GFP阳性细胞。