Serial viral passage and reverse genetics protocol for identifying the antiviral drug targets of EV-D68 antivirals

Haozhou Tan; Kan Li; Jun Wang

doi:10.1016/j.xpro.2025.104053

Serial viral passage and reverse genetics protocol for identifying the antiviral drug targets of EV-D68 antivirals

Haozhou Tan
, Kan Li
, Jun Wang

Research output: Contribution to journal › Article › peer-review

Abstract

Identifying antiviral targets and mechanisms is crucial for drug discovery. Here, we present a protocol to select EV-D68 variants resistant to Jun6504 by combining serial passage and reverse genetics. We describe steps for viral serial passage, sequencing, and viral genome mutagenesis. We then detail procedures for producing and characterizing recombinant viruses. This protocol provides a robust strategy to identify targets from phenotypic screening hits.

Original language	English (US)
Article number	104053
Journal	STAR Protocols
Volume	6
Issue number	3
DOIs	https://doi.org/10.1016/j.xpro.2025.104053
State	Published - Sep 19 2025
Externally published	Yes

Keywords

Chemistry
Microbiology
Molecular Biology

ASJC Scopus subject areas

General Neuroscience
General Immunology and Microbiology
General Biochemistry, Genetics and Molecular Biology

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10.1016/j.xpro.2025.104053

Cite this

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abstract = "Identifying antiviral targets and mechanisms is crucial for drug discovery. Here, we present a protocol to select EV-D68 variants resistant to Jun6504 by combining serial passage and reverse genetics. We describe steps for viral serial passage, sequencing, and viral genome mutagenesis. We then detail procedures for producing and characterizing recombinant viruses. This protocol provides a robust strategy to identify targets from phenotypic screening hits.",

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N2 - Identifying antiviral targets and mechanisms is crucial for drug discovery. Here, we present a protocol to select EV-D68 variants resistant to Jun6504 by combining serial passage and reverse genetics. We describe steps for viral serial passage, sequencing, and viral genome mutagenesis. We then detail procedures for producing and characterizing recombinant viruses. This protocol provides a robust strategy to identify targets from phenotypic screening hits.

AB - Identifying antiviral targets and mechanisms is crucial for drug discovery. Here, we present a protocol to select EV-D68 variants resistant to Jun6504 by combining serial passage and reverse genetics. We describe steps for viral serial passage, sequencing, and viral genome mutagenesis. We then detail procedures for producing and characterizing recombinant viruses. This protocol provides a robust strategy to identify targets from phenotypic screening hits.

KW - Chemistry

KW - Microbiology

KW - Molecular Biology

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Serial viral passage and reverse genetics protocol for identifying the antiviral drug targets of EV-D68 antivirals

Abstract

Keywords

ASJC Scopus subject areas

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