Generation of Recombinant SARS-CoV-2 Using a Bacterial Artificial Chromosome.

Generation of Recombinant SARS-CoV-2 Using a Bacterial Artificial Chromosome.
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DOI:
10.1002/cpmc.126
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发表时间:
2020-12-01
影响因子:
--
通讯作者:
Martinez-Sobrido, Luis
Martinez-Sobrido, Luis
中科院分区:
其他
文献类型:
--
作者:
Chiem, Kevin;Ye, Chengjin;Martinez-Sobrido, Luis

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截至2020年9月,SARS-CoV-2是COVID-19的病原体,已导致全球100万人死亡。在撰写本文时,尚无美国FDA批准的用于治疗SARS-CoV-2感染的治疗药物。在这里,我们描述了一个详细的协议,以产生重组(r)SARS-CoV-2使用反向遗传学方法的基础上使用的细菌人工染色体(BAC)。这种方法将允许生产突变的rSARS-CoV-2-这是必要的了解病毒蛋白质的功能,病毒的发病机制和/或传播,以及在病毒宿主界面的相互作用-和减毒的SARS-CoV-2,以促进发现有效的对策,以控制正在进行的SARS-CoV-2大流行。© 2020 Wiley Periodicals LLC.基本方案:使用细菌人工染色体生成重组SARS-CoV-2支持方案:rSARS-CoV-2的验证和表征。
SARS-CoV-2, the causative agent of COVID-19, has been responsible for a million deaths worldwide as of September 2020. At the time of this writing, there are no available US FDA-approved therapeutics for the treatment of SARS-CoV-2 infection. Here, we describe a detailed protocol to generate recombinant (r)SARS-CoV-2 using reverse-genetics approaches based on the use of a bacterial artificial chromosome (BAC). This method will allow the production of mutant rSARS-CoV-2-which is necessary for understanding the function of viral proteins, viral pathogenesis and/or transmission, and interactions at the virus-host interface-and attenuated SARS-CoV-2 to facilitate the discovery of effective countermeasures to control the ongoing SARS-CoV-2 pandemic. © 2020 Wiley Periodicals LLC. Basic Protocol: Generation of recombinant SARS-CoV-2 using a bacterial artificial chromosome Support Protocol: Validation and characterization of rSARS-CoV-2.