Influenza A virus (IAV) represents a significant burden on global health. This work proposes to generate a library of IAV strains each encoding a siRNA targeting known antiviral host genes with the goal of defining the potency of host factors in the context of an in vivo infection. For these studies, we will use a mouse-adapted influenza A virus that lacks a fully functional NS1 protein, thereby rendering the virus significantly attenuated (mIAV).
The specific aims of this proposal are to (1) engineer a mIAV-based siRNA library, (2) perform an in vivo RNAi screen to identify host determinants of IAV replication (3) to characterize the identified host restriction factors. To complete this work, a library of siRNAs was introduced into our mIAV, and the viruses were rescued individually. The viruses were used to infect mice to determine which viruses dominated in the population. Several known antiviral factors were identified in a preliminary screen that includes Trim21, Tlr7, RnaseL, Tbk1 and Irf1. In addition, two factors (Zfp182 and Mapk8) with unknown antiviral activity were identified which are currently being characterized.

Public Health Relevance

This project seeks to identify the host factors that restrict Influenza A virus (IAV) infection in vivo. IAV remains a significant health concern and identifying these host restriction factors is therefore critically important to elucidating how these factors restrict virus replication in vivo.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Predoctoral Individual National Research Service Award (F31)
Project #
1F31AI110130-01A1
Application #
8785890
Study Section
Special Emphasis Panel (ZRG1)
Program Officer
Adger-Johnson, Diane S
Project Start
2014-08-01
Project End
2017-07-31
Budget Start
2014-08-01
Budget End
2015-07-31
Support Year
1
Fiscal Year
2014
Total Cost
Indirect Cost
Name
Icahn School of Medicine at Mount Sinai
Department
Microbiology/Immun/Virology
Type
Schools of Medicine
DUNS #
City
New York
State
NY
Country
United States
Zip Code
10029
Benitez, Asiel A; Panis, Maryline; Xue, Jia et al. (2015) In Vivo RNAi Screening Identifies MDA5 as a Significant Contributor to the Cellular Defense against Influenza A Virus. Cell Rep 11:1714-26
Benitez, Asiel Arturo; Spanko, Laura Adrienne; Bouhaddou, Mehdi et al. (2015) Engineered Mammalian RNAi Can Elicit Antiviral Protection that Negates the Requirement for the Interferon Response. Cell Rep 13:1456-66