Identification of Required Host Factors for SARS-CoV-2 Infection in Human Cells.
Identification of Required Host Factors for SARS-CoV-2 Infection in Human Cells.
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DOI：
10.1016/j.cell.2020.10.030
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发表时间：
2021-01-07
期刊：
影响因子：
64.5
通讯作者：
Sanjana NE
中科院分区：
文献类型：
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作者：
Daniloski Z;Jordan TX;Wessels HH;Hoagland DA;Kasela S;Legut M;Maniatis S;Mimitou EP;Lu L;Geller E;Danziger O;Rosenberg BR;Phatnani H;Smibert P;Lappalainen T;tenOever BR;Sanjana NE
为了更好地了解宿主-病毒遗传依赖性并找到COVID-19的潜在治疗靶点，我们进行了基因组规模的CRISPR功能丧失筛选，以确定SARS-CoV-2病毒感染人肺泡上皮细胞所需的宿主因子。排名靠前的基因聚集成不同的通路，包括液泡ATP酶质子泵、Retromer和Commander复合物。我们使用几种正交方法验证这些基因靶点，如CRISPR敲除，RNA干扰敲除和小分子抑制剂。使用单细胞RNA测序，我们确定了胆固醇生物合成中的共享转录变化后的损失排名靠前的基因。此外，鉴于ACE 2受体在病毒进入的早期阶段的关键作用，我们表明，RAB 7A的损失减少病毒进入细胞内的隔离ACE 2受体。总的来说，这项工作提供了一个基因组规模的，定量资源的影响，每个宿主基因的损失对健身/响应病毒感染。为了确定SARS-CoV-2的潜在治疗靶点，Daniloski et al.在人类肺上皮细胞中进行全基因组CRISPR筛选。他们确定了SARS-CoV-2感染所需的基因和途径，包括液泡ATP酶质子泵，Retromer和Commander复合物。使用单细胞转录组学，他们确定胆固醇生物合成的上调是病毒耐药性的常见机制，除了ACE 2隔离。
To better understand host-virus genetic dependencies and find potential therapeutic targets for COVID-19, we performed a genome-scale CRISPR loss-of-function screen to identify host factors required for SARS-CoV-2 viral infection of human alveolar epithelial cells. Top-ranked genes cluster into distinct pathways, including the vacuolar ATPase proton pump, Retromer, and Commander complexes. We validate these gene targets using several orthogonal methods such as CRISPR knockout, RNA interference knockdown, and small-molecule inhibitors. Using single-cell RNA-sequencing, we identify shared transcriptional changes in cholesterol biosynthesis upon loss of top-ranked genes. In addition, given the key role of the ACE2 receptor in the early stages of viral entry, we show that loss of RAB7A reduces viral entry by sequestering the ACE2 receptor inside cells. Overall, this work provides a genome-scale, quantitative resource of the impact of the loss of each host gene on fitness/response to viral infection. To identify potential therapeutic targets for SARS-CoV-2, Daniloski et al. conduct a genome-wide CRISPR screen in human lung epithelial cells. They identify genes and pathways required for SARS-CoV-2 infection, including the vacuolar ATPase proton pump, Retromer, and Commander complexes. Using single-cell transcriptomics, they identify upregulation of cholesterol biosynthesis as a common mechanism underlying viral resistance, in addition to ACE2 sequestration.
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影响因子：
14.9
作者：
Kuleshov MV;Jones MR;Rouillard AD;Fernandez NF;Duan Q;Wang Z;Koplev S;Jenkins SL;Jagodnik KM;Lachmann A;McDermott MG;Monteiro CD;Gundersen GW;Ma'ayan A
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影响因子：
64.5
作者：
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通讯作者：
Krogan, Nevan J.
影响因子：
6
作者：
Guerra F;Bucci C
通讯作者：
Bucci C
DOI：
10.1038/nrmicro2090
发表时间：
2009-03
期刊：
Nature reviews. Microbiology
影响因子：
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作者：
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