严重急性呼吸综合征冠状病毒2（SARS-CoV-2）是导致2019年冠状病毒病的病原体，已成为全球关注的健康问题。因此，有一个巨大的需求通过使用与人类疾病相关的细胞来了解病毒与宿主的相互作用网络。因此，我们使用单倍体人类胚胎干细胞（hESCs）进行了一个失活性基因组范围的筛选，以鉴定参与SARS-CoV-2感染的基因。尽管未分化的hESCs对SARS-CoV-2具有抵抗性，但其高水平表达ACE2的分化出的定义性内胚层（DE）后代却对该病毒高度敏感。我们的基因筛选能够确认良好建立的入侵受体ACE2作为宿主因子，并且还发现了潜在的SARS-CoV-2新型调控因子。其中两个新型筛选命中物，转录因子MAFG和跨膜蛋白TMEM86A，通过CRISPR介导的hESCs突变验证进一步证实能够使DE细胞对SARS-CoV-2具有抵抗力，在突变株分化为DE细胞并接种SARS-CoV-2感染后进行了验证。我们的基因组范围遗传筛查研究了非癌人类细胞中的SARS-CoV-2宿主因子，这些细胞具有内源性ACE2表达，为在人类细胞中识别SARS-CoV-2细胞病理学新型调控因子提供了独特平台。版权© 2023年作者。由Elsevier公司出版。保留所有权利。

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), the causative agent of coronavirus disease 2019, has become a global health concern. Therefore, there is an immense need to understand the network of virus-host interactions by using human disease-relevant cells. We have thus conducted a loss-of-function genome-wide screen using haploid human embryonic stem cells (hESCs) to identify genes involved in SARS-CoV-2 infection. Although the undifferentiated hESCs are resistant to SARS-CoV-2, their differentiated definitive endoderm (DE) progenies, which express high levels of ACE2, are highly sensitive to the virus. Our genetic screening was able to identify the well-established entry receptor ACE2 as a host factor, along with additional potential novel modulators of SARS-CoV-2. Two such novel screen hits, the transcription factor MAFG and the transmembrane protein TMEM86A, were further validated as conferring resistance against SARS-CoV-2 by using CRISPR-mediated mutagenesis in hESCs, followed by differentiation of mutant lines into DE cells and infection by SARS-CoV-2. Our genome-wide genetic screening investigated SARS-CoV-2 host factors in non-cancerous human cells with endogenous ACE2 expression, providing a unique platform to identify novel modulators of SARS-CoV-2 cytopathology in human cells.Copyright © 2023 The Author(s). Published by Elsevier Inc. All rights reserved.