Event Title

Generation of Influenza NP-FLAG Virus to Facilitate NP Interaction Studies

Presenter Information

Joscelyn Berumen
Raquel Rodriguez

Presentation Type

Poster Presentation

College

College of Natural Sciences

Location

SMSU Event Center BC

Faculty Mentor

Dr. Laura Newcomb

Start Date

5-16-2019 9:30 AM

End Date

5-16-2019 11:00 AM

Abstract

Influenza viruses cause seasonal epidemics of respiratory illness. Influenza is combated with annual vaccinations due to the rapidly changing nature of the virus, antiviral therapies become ineffective emphasizing need for new antiviral targets. The influenza viral ribonucleoprotein (vRNP) is responsible for influenza RNA synthesis and is highly conserved across multiple influenza subtypes, making this an excellent antiviral target. Nucleoprotein (NP) is an essential component of vRNPs and interacts with both viral and host factors to regulate viral RNA expression. We aim to examine NP protein interactions during the viral life cycle through generation of influenza virus expressing NP-FLAG tagged protein. Here we constructed a DNA plasmid to express NP-FLAG coding sequence with an additional repeat of RNA containing packaging signals (PS) reported to result in 60% packaging efficiency. virus. Samples were collected tested for virion presence in hemagglutination assay (HA). While we obtained WT-NP virions, we did not detect virions from NP-FLAG-PS. We will attempt to improve WT-NP virus yield to recover NP-FLAG-PS virus. If successful, generation of NP-FLAG influenza virus will provide a tool for the influenza community to study NP and NP interactions.

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May 16th, 9:30 AM May 16th, 11:00 AM

Generation of Influenza NP-FLAG Virus to Facilitate NP Interaction Studies

SMSU Event Center BC

Influenza viruses cause seasonal epidemics of respiratory illness. Influenza is combated with annual vaccinations due to the rapidly changing nature of the virus, antiviral therapies become ineffective emphasizing need for new antiviral targets. The influenza viral ribonucleoprotein (vRNP) is responsible for influenza RNA synthesis and is highly conserved across multiple influenza subtypes, making this an excellent antiviral target. Nucleoprotein (NP) is an essential component of vRNPs and interacts with both viral and host factors to regulate viral RNA expression. We aim to examine NP protein interactions during the viral life cycle through generation of influenza virus expressing NP-FLAG tagged protein. Here we constructed a DNA plasmid to express NP-FLAG coding sequence with an additional repeat of RNA containing packaging signals (PS) reported to result in 60% packaging efficiency. virus. Samples were collected tested for virion presence in hemagglutination assay (HA). While we obtained WT-NP virions, we did not detect virions from NP-FLAG-PS. We will attempt to improve WT-NP virus yield to recover NP-FLAG-PS virus. If successful, generation of NP-FLAG influenza virus will provide a tool for the influenza community to study NP and NP interactions.