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Multiwell plate shaker

Company: Thermo Fisher Scientific
Catalog#: 75004524
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Retroviral Capsid Core Stability Assay
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2018-09-20
[Abstract]  Structural stability of the capsid core is a critical parameter for the productive infection of a cell by a retrovirus. Compromised stability can lead to premature core disassembly, exposure of replication intermediates to cytosolic nucleic acid sensors that can trigger innate antiviral responses, and failure to integrate the proviral genome into the host DNA. Thus, core stability is a critical feature of viral replicative fitness. While there are several well-described techniques to assess viral capsid core stability, most are generally time and labor intensive. Recently, our group compared the relative stability of murine leukemia virus capsid cores using an in vitro detergent-based approach combined with ultracentrifugation against the popular fate of capsid assay. We found ... [摘要]  衣壳核心的结构稳定性是逆转录病毒对细胞的生产性感染的关键参数。受损的稳定性可导致核心过早解体,将复制中间体暴露于细胞溶质核酸传感器,其可触发先天的抗病毒反应,并且不能将原病毒基因组整合到宿主DNA中。因此,核心稳定性是病毒复制适应性的关键特征。虽然有几种充分描述的技术来评估病毒衣壳核心稳定性,但大多数通常是时间和劳动密集型的。最近,我们小组使用基于体外洗涤剂的方法结合超速离心法对衣壳测定的流行命运比较了鼠白血病病毒衣壳核心的相对稳定性。我们发现两种方法都得出了类似的结论,尽管第一种方法是比较显示核心稳定性差异的病毒突变体时评估相对衣壳核心稳定性的一种非常简单和快速的方法。

【背景】逆转录病毒已经进化出复制周期,在规避宿主抗病毒反应方面表现优异。逆转录病毒开发的一种策略是将其复制中间体与胞质核酸传感器如cGAS,TREX1,IFI203和DDX41屏蔽起来(Yan et al。,2010; Gao et al。,2013; Lahaye et al。,2013; Stavrou et al。,2015)。在复制过程中,逆转录病毒在细胞质中产生RNA-DNA杂合体和未甲基化的双链前病毒DNA,这是先天免疫传感器的常见靶标(Yan et al。,2010; Gao et al。,2013; Lahaye et ...

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