Construction and Transcriptomic Study of Chicken IFNAR1-Knockout Cell Line Reveals the Essential Roles of Cell Growth- and Apoptosis-Related Pathways in Duck Tembusu Virus Infection

Viruses. 2022 Oct 9;14(10):2225. doi: 10.3390/v14102225.

Abstract

For industrial vaccine production, overwhelming the existing antiviral innate immune response dominated by type I interferons (IFN-I) in cells would be a key factor improving the effectiveness and production cost of vaccines. In this study, we report the construction of an IFN-I receptor 1 (IFNAR1)-knockout DF-1 cell line (KO-IFNAR1), which supports much more efficient replication of the duck Tembusu virus (DTMUV), Newcastle disease virus (NDV) and gammacoronavirus infectious bronchitis virus (IBV). Transcriptomic analysis of DTMUV-infected KO-IFNAR1 cells demonstrated that DTMUV mainly activated genes and signaling pathways related to cell growth and apoptosis. Among them, JUN, MYC and NFKBIA were significantly up-regulated. Furthermore, knockdown of zinc-fingered helicase 2 (HELZ2) and interferon-α-inducible protein 6 (IFI6), the two genes up-regulated in both wild type and KO-IFNAR1 cells, significantly increased the replication of DTMUV RNA. This study paves the way for further studying the mechanism underlying the DTMUV-mediated IFN-I-independent regulation of virus replication, and meanwhile provides a potential cell resource for efficient production of cell-based avian virus vaccines.

Keywords: CRISPR/Cas9; DF-1; IFNAR1; duck Tembusu virus; transcriptomic analysis.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antiviral Agents
  • Apoptosis
  • Cell Line
  • Chickens / genetics
  • Ducks
  • Flavivirus Infections*
  • Flavivirus* / genetics
  • Interferon Type I* / genetics
  • Interferon-alpha / genetics
  • Poultry Diseases*
  • RNA
  • Transcriptome
  • Zinc

Substances

  • Interferon Type I
  • Antiviral Agents
  • RNA
  • Interferon-alpha
  • Zinc

Grants and funding

This research was funded by the Independent project of Zhaoqing Branch Center of Guangdong Laboratory for Lingnan Modern Agricultural Science and Technology (No. P20211154-0301 and No. P20211154-0202) the Zhaoqing Science Foundation of China (Grant Nos. ZK. 2019K040 and ZK. 2020NCP0102), and Guangdong Province Key Field R&D Program (No. 2021B0707010009).