title: Replication-competent influenza A viruses expressing a red fluorescent protein.
abstract: Like most animal viruses, studying influenza A in model systems requires secondary methodologies to identify infected cells. To circumvent this requirement, we describe the generation of replication-competent influenza A red fluorescent viruses. These influenza A viruses encode mCherry fused to the viral non-structural 1 (NS1) protein and display comparable growth kinetics to wild-type viruses in vitro. Infection of cells with influenza A mCherry viruses was neutralized with monoclonal antibodies and inhibited with antivirals to levels similar to wild-type virus. Influenza A mCherry viruses were also able to lethally infect mice, and strikingly, dose- and time-dependent kinetics of viral replication were monitored in whole excised mouse lungs using an in vivo imaging system (IVIS). By eliminating the need for secondary labeling of infected cells, influenza A mCherry viruses provide an ideal tool in the ongoing struggle to better characterize the virus and identify new therapeutics against influenza A viral infections.
authors.affiliation: Department of Microbiology and Immunology, University of Rochester, Rochester, NY, United States.
authors.forename: Aitor
authors.initials: A
authors.lastname: Nogales
authors.affiliation: Department of Microbiology and Immunology, University of Rochester, Rochester, NY, United States.
authors.forename: Steven F
authors.initials: SF
authors.lastname: Baker
authors.affiliation: Department of Microbiology and Immunology, University of Rochester, Rochester, NY, United States. Electronic address: luis_martinez@urmc.rochester.edu.
authors.forename: Luis
authors.initials: L
authors.lastname: Martínez-Sobrido
mesh_terms: D000818:Animals; D002460:Cell Line; D058948:Cell Tracking / Q000379:methods*; D005260:Female; D015870:Gene Expression; D006801:Humans; D053118:Influenza A Virus, H1N1 Subtype / Q000235:genetics / Q000502:physiology*; D009980:Influenza A virus / Q000235:genetics / Q000502:physiology*; D007251:Influenza, Human / Q000821:virology*; D008164:Luminescent Proteins / Q000235:genetics* / Q000378:metabolism; D051379:Mice; D008810:Mice, Inbred C57BL; D017361:Viral Nonstructural Proteins / Q000235:genetics / Q000378:metabolism; D014779:Virus Replication; D000097573:Red Fluorescent Protein
chemical_list: D008164:Luminescent Proteins; D017361:Viral Nonstructural Proteins; D000097573:Red Fluorescent Protein; C085820:INS1 protein, influenza virus
keywords: 2A; Antivirals; In vivo imaging system (IVIS); Influenza A virus; Interferon; Microneutralization assay; NS1; Neutralizing antibodies; Replication-competent virus; Virus neutralization assay; mCherry