Expression of a single siRNA against a conserved region of NP gene strongly inhibits in vitro replication of different Influenza A virus strains of avian and swine origin

Elena Stoppani, Ivan Bassi, Silvia Dotti, Michela Lizier, Maura Ferrari, Franco Lucchini

Research output: Contribution to journalArticle

Abstract

Influenza A virus is the principal agent responsible of the respiratory tract's infections in humans. Every year, highly pathogenic and infectious strains with new antigenic assets appear, making ineffective vaccines so far developed. The discovery of RNA interference (RNAi) opened the way to the progress of new promising drugs against Influenza A virus and also to the introduction of disease resistance traits in genetically modified animals. In this paper, we show that Madin-Darby Canine Kidney (MDCK) cell line expressing short hairpin RNAs (shRNAs) cassette, designed on a specific conserved region of the nucleoprotein (NP) viral genome, can strongly inhibit the viral replication of four viral strains sharing the target sequence, reducing the viral mRNA respectively to 2.5 × 10-4, 7.5 × 10-5, 1.7 × 10-3, 1.9 × 10-4 compared to the control, as assessed by real-time PCR. Moreover, we demonstrate that during the challenge with a viral strain bearing a single mismatch on the target sequence, although a weaker inhibition is observed, viral mRNA is still lowered down to 1.2 × 10-3 folds in the shRNA-expressing clone compared to the control, indicating a broad potential use of this approach. In addition, we developed a highly predictive and fast screening test of siRNA sequences based on dual-luciferase assay, useful for the in vitro prediction of the potential effect of viral inhibition. In conclusion, these findings reveal new siRNA sequences able to inhibit Influenza A virus replication and provide a basis for the development of siRNAs as prophylaxis and therapy for influenza infection both in humans and animals.

Original languageEnglish
Pages (from-to)16-22
Number of pages7
JournalAntiviral Research
Volume120
DOIs
Publication statusPublished - Aug 1 2015

Fingerprint

Nucleoproteins
Influenza A virus
Small Interfering RNA
Swine
Genes
Messenger RNA
Genetically Modified Animals
Madin Darby Canine Kidney Cells
Disease Resistance
Viral Genome
Virus Replication
RNA Interference
Luciferases
Respiratory Tract Infections
Human Influenza
Real-Time Polymerase Chain Reaction
Vaccines
Clone Cells
Cell Line
In Vitro Techniques

Keywords

  • Influenza
  • Nucleoprotein
  • Orthomyxoviridae
  • RNA interference
  • siRNA

ASJC Scopus subject areas

  • Virology
  • Pharmacology

Cite this

Expression of a single siRNA against a conserved region of NP gene strongly inhibits in vitro replication of different Influenza A virus strains of avian and swine origin. / Stoppani, Elena; Bassi, Ivan; Dotti, Silvia; Lizier, Michela; Ferrari, Maura; Lucchini, Franco.

In: Antiviral Research, Vol. 120, 01.08.2015, p. 16-22.

Research output: Contribution to journalArticle

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