Live Imaging and Quantification of Viral Infection in K18 hACE2 Transgenic Mice Using Reporter-Expressing Recombinant SARS-CoV-2.

Live Imaging and Quantification of Viral Infection in K18 hACE2 Transgenic Mice Using Reporter-Expressing Recombinant SARS-CoV-2.
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DOI:
10.3791/63127
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发表时间:
2021-11-05
期刊:
Journal of visualized experiments : JoVE
影响因子:
--
通讯作者:
Martínez-Sobrido L
Martínez-Sobrido L
中科院分区:
其他
文献类型:
--
作者:
Morales Vasquez D;Chiem K;Silvas J;Park JG;Ye C;Martínez-Sobrido L

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2019冠状病毒病(COVID-19)大流行是由严重急性呼吸综合征冠状病毒2 (SARS-CoV-2)引起的。迄今为止,SARS-CoV-2已在全球造成超过2.42亿人感染,490多万人死亡。与其他病毒类似,研究SARS-CoV-2需要使用实验方法来检测受感染细胞和/或动物模型中是否存在病毒。为了克服这一限制,我们产生了具有复制能力的重组(r)SARS-CoV-2,表达生物发光(纳米荧光素酶,Nluc)或荧光(金星)蛋白。基于Nluc和Venus报告基因的表达,这些表达报告基因的sars - cov -2可以在体外和体内追踪病毒感染。本研究描述了使用SARS-CoV-2/Nluc和SARS-CoV-2/Venus在先前描述的K18人血管紧张素转换酶2 (hACE2)转基因小鼠感染模型中使用体内成像系统(IVIS)检测和跟踪SARS-CoV-2感染。该rssars - cov -2/Nluc和rssars - cov -2/Venus显示rssars - cov -2/ wt样致病性和病毒在体内复制。重要的是,Nluc和Venus的表达使我们能够在感染小鼠体内和体外直接跟踪病毒感染。这些SARS-CoV-2/Nluc和SARS-CoV-2/Venus是研究SARS-CoV-2体内生物学,了解病毒感染和相关的COVID-19疾病,以及确定有效的预防和/或治疗方法以对抗SARS-CoV-2感染的绝佳选择。
The coronavirus disease 2019 (COVID-19) pandemic has been caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). To date, SARS-CoV-2 has been responsible for over 242 million infections and more than 4.9 million deaths worldwide. Similar to other viruses, studying SARS-CoV-2 requires the use of experimental methods to detect the presence of virus in infected cells and/or in animal models. To overcome this limitation, we generated replication-competent recombinant (r)SARS-CoV-2 that expresses bioluminescent (nanoluciferase, Nluc) or fluorescent (Venus) proteins. These reporter-expressing rSARS-CoV-2 allow tracking viral infections in vitro and in vivo based on the expression of Nluc and Venus reporter genes. Here the study describes the use of rSARS-CoV-2/Nluc and rSARS-CoV-2/Venus to detect and track SARS-CoV-2 infection in the previously described K18 human angiotensin-converting enzyme 2 (hACE2) transgenic mouse model of infection using in vivo imaging systems (IVIS). This rSARS-CoV-2/Nluc and rSARS-CoV-2/Venus show rSARS-CoV-2/WT-like pathogenicity and viral replication in vivo. Importantly, Nluc and Venus expression allow us to directly track viral infections in vivo and ex vivo, in infected mice. These rSARS-CoV-2/Nluc and rSARS-CoV-2/Venus represent an excellent option to study the biology of SARS-CoV-2 in vivo, to understand viral infection and associated COVID-19 disease, and to identify effective prophylactic and/or therapeutic treatments to combat SARS-CoV-2 infection.
BNT162B2 mRNA COVID-19疫苗的安全性和功效。
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影响因子: --
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影响因子: --
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DOI: 10.7759/cureus.15280
发表时间: 2021-05-27
期刊: Cureus
影响因子: --
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DOI: 10.1016/j.antiviral.2019.05.013
发表时间: 2019-08-01
期刊: ANTIVIRAL RESEARCH
影响因子: 7.6
作者:
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通讯作者: Zhang, Bo
DOI: 10.1016/j.jbc.2021.100847
发表时间: 2021-07
期刊: The Journal of biological chemistry
影响因子: --
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通讯作者: Liu SL