Construction and Rescue of a DNA-Launched DENV2 Infectious Clone.
Construction and Rescue of a DNA-Launched DENV2 Infectious Clone.
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DOI：
10.3390/v15020275
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发表时间：
2023-01-18
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影响因子：
--
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黄病毒代表了一大组全球重要的昆虫传播病原体。对于这些病毒中的许多病毒，缺乏抗病毒药物和疫苗。因此，有必要继续开发工具，包括反向遗传技术，以进一步推动我们防治这些病原体的努力。传统上，黄病毒的反向遗传学方法依赖于从体外转录反应产生感染性RNA，然后电穿孔或转染到允许的细胞系中。然而，从CMV启动子驱动的表达质粒生产寨卡病毒已经成功，这提供了成本和时间优势。在这份报告中，我们描述了登革病毒（DENV）血清型2株16681的DNA启动的感染性克隆的设计和构建。在病毒基因组的非结构蛋白1区段中引入人工内含子以促进在细菌中的稳定性。我们发现，拯救的病毒保持了在常用细胞系中形成噬斑和有效复制的能力。因此，我们提出了一种从质粒DNA生产DENV 2菌株16681的快速且具有成本效益的方法。该构建体将成为持续开发抗DENV治疗剂和疫苗的有用平台。
Flaviviruses represent a large group of globally significant, insect-borne pathogens. For many of these viruses, there is a lack of antivirals and vaccines. Thus, there is a need to continue the development of tools to further advance our efforts to combat these pathogens, including reverse genetics techniques. Traditionally, reverse genetics methods for flaviviruses rely on producing infectious RNA from in vitro transcription reactions followed by electroporation or transfection into permissive cell lines. However, the production of Zika virus has been successful from CMV promoter-driven expression plasmids, which provides cost and time advantages. In this report, we describe the design and construction of a DNA-launched infectious clone for dengue virus (DENV) serotype 2 strain 16681. An artificial intron was introduced in the nonstructural protein 1 segment of the viral genome to promote stability in bacteria. We found that rescued viruses maintained the ability to form plaques and replicate efficiently in commonly used cell lines. Thus, we present a rapid and cost-effective method for producing DENV2 strain 16681 from plasmid DNA. This construct will be a useful platform for the continued development of anti-DENV therapeutics and vaccines.
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影响因子：
4.8
作者：
Schwarz MC;Sourisseau M;Espino MM;Gray ES;Chambers MT;Tortorella D;Evans MJ
通讯作者：
Evans MJ
影响因子：
158.5
作者：
Hadinegoro, S. R.;Arredondo-Garcia, J. L.;Saville, M.
通讯作者：
Saville, M.
DOI：
10.1073/pnas.88.12.5139
发表时间：
1991-06-01
影响因子：
11.1
作者：
LAI, CJ;ZHAO, B;BRAY, M
通讯作者：
BRAY, M
影响因子：
3.7
作者：
Ansarah-Sobrinho, Camilo;Nelson, Steevenson;Jost, Christiane A.;Whitehead, Stephen S.;Pierson, Theodore C.
通讯作者：
Pierson, Theodore C.
影响因子：
3.8
作者：
Mandl, CW;Ecker, M;Heinz, FX
通讯作者：
Heinz, FX