Characterization of an RNA binding protein required for influenza virus replication

/ABSTRACT The overarching goal of this application is to define and investigate host factors required for influenza virus replication. Despite current interventions, influenza viruses are responsible for a significant portion of upper respiratory infections globally. This has prompted exploration of how these viruses interact with host cells to facilitate their replication. We conducted a series of genome-wide CRISPR screens with nine genetically divergent influenza viral strains to explore shared host-dependency factors. We identified one such host factor, TTC14, an uncharacterized RNA binding protein completely required for the replication of all influenza virus strains. The studies proposed in this application will provide important insights into the role this protein plays in regulating RNA processing and how its absence impacts influenza virus infection. The goals of this application are premised on the central hypothesis that TTC14 binds to transcripts and coordinates RNA splicing, leading to proper downstream protein translation. In Aim 1, I will investigate the molecular mechanism of TTC14-regulated mRNA splicing in the absence of and during influenza infection. In Aim 2, I will characterize how the loss of TTC14 expression impacts influenza virus replication in primary human airway cells and how this alters host immune responses against viral infection. Notably, this research will illuminate a novel regulatory mechanism of RNA splicing and reveal a new interface between influenza virus and host machinery that many strains manipulate to aid their replication. Given the extensive expertise of my lab in studying host factors involved in viral replication, I am uniquely positioned to perform these studies, which will provide new paradigms in characterizing the complex host-viral interactions of influenza viruses. Project Number: 1F30AI183629-01A1 | Fiscal Year: 2025 | NIH Institute/Center: National Institute of Allergy and Infectious Diseases (NIAID) | Principal Investigator: Vishwas Rao | Institution: DUKE UNIVERSITY, DURHAM, NC | Award Amount: $42,815 | Activity Code: F30 | Study Section: Special Emphasis Panel[ZRG1 F07C-H (20)] View on NIH RePORTER: https://reporter.nih.gov/project-details/1F30AI18362901A1