A plasmid-based reporter system for live cell imaging of dengue virus infected cells.

A plasmid-based reporter system for live cell imaging of dengue virus infected cells.
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DOI:
10.1016/j.jviromet.2014.10.010
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发表时间:
2015-01
影响因子:
3.1
通讯作者:
Rothman AL
Rothman AL
中科院分区:
医学4区
文献类型:
--
作者:
Medin CL;Valois S;Patkar CG;Rothman AL

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细胞培养模型广泛用于研究登革热病毒(DENV)对宿主细胞功能的影响。目前鉴定感染未修饰 DENV 的细胞的方法需要对细胞进行固定和透化,以进行 DENV 特异性抗体染色。该方法不允许随时间推移对活细胞进行成像。在本报告中,开发了一种基于质粒的报告基因,可以对感染 DENV 的细胞进行非破坏性鉴定。基于质粒的报告基因被证明广泛适用于四种 DENV 血清型,包括低传代毒株,并且可以被病毒蛋白酶特异性切割,对病毒生产的干扰最小。这项研究揭示了这种新型报告系统在推进 DENV 感染期间病毒与宿主相互作用的研究方面的潜力。
Cell culture models are used widely to study the effects of dengue virus (DENV) on host cell function. Current methods of identification of cells infected with an unmodified DENV requires fixation and permeablization of cells to allow DENV-specific antibody staining. This method does not permit imaging of viable cells over time. In this report, a plasmid-based reporter was developed to allow non-destructive identification of DENV-infected cells. The plasmid-based reporter was demonstrated to be broadly applicable to the four DENV serotypes, including low-passaged strains, and was specifically cleaved by the viral protease with minimal interference on viral production. This study reveals the potential for this novel reporter system to advance the studies of virus-host interactions during DENV infection.
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