High-throughput investigation of genetic design constraints in domesticated Influenza A Virus for transient gene delivery.

Autor:	Sychla A; Department of Biochemistry, Molecular Biology, and Biophysics, University of Minnesota, Saint Paul, MN 55108.; Biotechnology Institute, University of Minnesota, Saint Paul, MN 55108., Stach CS; Department of Biochemistry, Molecular Biology, and Biophysics, University of Minnesota, Saint Paul, MN 55108.; Biotechnology Institute, University of Minnesota, Saint Paul, MN 55108., Roach SN; Department of Biochemistry, Molecular Biology, and Biophysics, University of Minnesota, Saint Paul, MN 55108.; Department of Microbiology and Immunology, University of Minnesota, Saint Paul, MN 55108., Hayward AN; Department of Biochemistry, Molecular Biology, and Biophysics, University of Minnesota, Saint Paul, MN 55108.; Biotechnology Institute, University of Minnesota, Saint Paul, MN 55108., Langlois RA; Department of Biochemistry, Molecular Biology, and Biophysics, University of Minnesota, Saint Paul, MN 55108.; Department of Microbiology and Immunology, University of Minnesota, Saint Paul, MN 55108., Smanski MJ; Department of Biochemistry, Molecular Biology, and Biophysics, University of Minnesota, Saint Paul, MN 55108.; Biotechnology Institute, University of Minnesota, Saint Paul, MN 55108.
Jazyk:	angličtina
Zdroj:	BioRxiv : the preprint server for biology [bioRxiv] 2024 Feb 15. Date of Electronic Publication: 2024 Feb 15.
DOI:	10.1101/2024.02.14.580300
Abstrakt:	Replication-incompetent single cycle infectious Influenza A Virus (sciIAV) has demonstrated utility as a research and vaccination platform. Protein-based therapeutics are increasingly attractive due to their high selectivity and potent efficacy but still suffer from low bioavailability and high manufacturing cost. Transient RNA-mediated delivery is a safe alternative that allows for expression of protein-based therapeutics within the target cells or tissues but is limited by delivery efficiency. Here, we develop recombinant sciIAV as a platform for transient gene delivery in vivo and in vitro for therapeutic, research, and manufacturing applications ( in vivo antimicrobial production, cell culture contamination clearance, and production of antiviral proteins in vitro ). While adapting the system to deliver new protein cargo we discovered expression differences presumably resulting from genetic context effects. We applied a high-throughput screen to map these within the 3'-untranslated and coding regions of the hemagglutinin-encoding segment 4. This screen revealed permissible mutations in the 3'-UTR and depletion of RNA level motifs in the N-terminal coding region.
Databáze:	MEDLINE
Externí odkaz:	Zobrazit plný text záznamu