A CRISPR/Cas9 genetically engineered organoid biobank reveals essential host factors for coronaviruses.
A CRISPR/Cas9 genetically engineered organoid biobank reveals essential host factors for coronaviruses.
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DOI：
10.1038/s41467-021-25729-7
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发表时间：
2021-09-17
影响因子：
16.6
通讯作者：
Clevers H
中科院分区：
文献类型：
--
作者：
Beumer J;Geurts MH;Lamers MM;Puschhof J;Zhang J;van der Vaart J;Mykytyn AZ;Breugem TI;Riesebosch S;Schipper D;van den Doel PB;de Lau W;Pleguezuelos-Manzano C;Busslinger G;Haagmans BL;Clevers H
快速鉴定病毒复制所必需的宿主基因可能会加快治疗干预措施的产生。遗传筛选通常在转化的细胞系中进行，这些细胞系在体内不能很好地代表病毒靶细胞，从而导致可能无法转化为临床的发现。肠道类器官越来越多地用于模拟人类疾病，并适用于基因工程。为了辨别哪些宿主因子是可靠的抗冠状病毒治疗靶点，我们为先前与冠状病毒生物学有关的19个宿主基因生成突变克隆IO。我们验证ACE 2和DPP 4分别是SARS-CoV/SARS-CoV-2和MERS-CoV的进入受体。IOs中的SARS-CoV-2复制不需要内体组织蛋白酶B/L蛋白酶，但特别依赖于细胞表面蛋白酶TMPRSS 2。其他TMPRSS家族成员不是必需的。新出现的冠状病毒变体B.1.1.7以及SARS-CoV和MERS-CoV同样依赖于TMPRSS 2。这些发现强调了非转化人类模型与冠状病毒研究的相关性，将TMPRSS 2确定为有吸引力的泛冠状病毒治疗靶点，并证明类器官敲除生物库是研究当前和未来新兴冠状病毒生物学的有价值工具。快速鉴定病毒复制所必需的宿主基因可能会加快治疗干预措施的产生。在这里，作者为先前与冠状病毒生物学有关的19个宿主基因生成了突变克隆肠道类器官，并将细胞表面蛋白酶TMPRSS 2确定为潜在的治疗靶点。
Rapid identification of host genes essential for virus replication may expedite the generation of therapeutic interventions. Genetic screens are often performed in transformed cell lines that poorly represent viral target cells in vivo, leading to discoveries that may not be translated to the clinic. Intestinal organoids are increasingly used to model human disease and are amenable to genetic engineering. To discern which host factors are reliable anti-coronavirus therapeutic targets, we generate mutant clonal IOs for 19 host genes previously implicated in coronavirus biology. We verify ACE2 and DPP4 as entry receptors for SARS-CoV/SARS-CoV-2 and MERS-CoV respectively. SARS-CoV-2 replication in IOs does not require the endosomal Cathepsin B/L proteases, but specifically depends on the cell surface protease TMPRSS2. Other TMPRSS family members were not essential. The newly emerging coronavirus variant B.1.1.7, as well as SARS-CoV and MERS-CoV similarly depended on TMPRSS2. These findings underscore the relevance of non-transformed human models for coronavirus research, identify TMPRSS2 as an attractive pan-coronavirus therapeutic target, and demonstrate that an organoid knockout biobank is a valuable tool to investigate the biology of current and future emerging coronaviruses. Rapid identification of host genes essential for virus replication may expedite the generation of therapeutic interventions. Here the authors generate mutant clonal intestinal organoids for 19 host genes previously implicated in coronavirus biology and identify the cell surface protease TMPRSS2 as a potential therapeutic target.
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DOI：
10.1126/science.abd3072
发表时间：
2020-11-13
期刊：
Science (New York, N.Y.)
影响因子：
--
作者：
Daly JL;Simonetti B;Klein K;Chen KE;Williamson MK;Antón-Plágaro C;Shoemark DK;Simón-Gracia L;Bauer M;Hollandi R;Greber UF;Horvath P;Sessions RB;Helenius A;Hiscox JA;Teesalu T;Matthews DA;Davidson AD;Collins BM;Cullen PJ;Yamauchi Y
通讯作者：
Yamauchi Y
影响因子：
64.5
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
通讯作者：
Sanjana NE
影响因子：
--
作者：
Hulswit RJ;de Haan CA;Bosch BJ
通讯作者：
Bosch BJ
DOI：
10.1016/s0140-6736(03)13967-0
发表时间：
2003-07-26
期刊：
Lancet (London, England)
影响因子：
--
作者：
Kuiken T;Fouchier RA;Schutten M;Rimmelzwaan GF;van Amerongen G;van Riel D;Laman JD;de Jong T;van Doornum G;Lim W;Ling AE;Chan PK;Tam JS;Zambon MC;Gopal R;Drosten C;van der Werf S;Escriou N;Manuguerra JC;Stöhr K;Peiris JS;Osterhaus AD
通讯作者：
Osterhaus AD
影响因子：
11.8
作者：
Kaye M
通讯作者：
Kaye M