Frontiers in Microbiology (Mar 2023)

An efficient double-fluorescence approach for generating fiber-2-edited recombinant serotype 4 fowl adenovirus expressing foreign gene

  • Yiwen Guo,
  • Yiwen Guo,
  • Yiwen Guo,
  • Yiwen Guo,
  • Zhenqi Xu,
  • Zhenqi Xu,
  • Zhenqi Xu,
  • Zhenqi Xu,
  • Yifei Chao,
  • Yifei Chao,
  • Yifei Chao,
  • Yifei Chao,
  • Xudong Cao,
  • Xudong Cao,
  • Xudong Cao,
  • Xudong Cao,
  • Huiru Jiang,
  • Huiru Jiang,
  • Huiru Jiang,
  • Huiru Jiang,
  • Han Li,
  • Han Li,
  • Han Li,
  • Han Li,
  • Tuofan Li,
  • Tuofan Li,
  • Tuofan Li,
  • Tuofan Li,
  • Zhimin Wan,
  • Zhimin Wan,
  • Zhimin Wan,
  • Zhimin Wan,
  • Hongxia Shao,
  • Hongxia Shao,
  • Hongxia Shao,
  • Hongxia Shao,
  • Aijian Qin,
  • Aijian Qin,
  • Aijian Qin,
  • Aijian Qin,
  • Quan Xie,
  • Quan Xie,
  • Quan Xie,
  • Quan Xie,
  • Jianqiang Ye,
  • Jianqiang Ye,
  • Jianqiang Ye,
  • Jianqiang Ye

DOI
https://doi.org/10.3389/fmicb.2023.1160031
Journal volume & issue
Vol. 14

Abstract

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Recently, the infection of serotype 4 fowl adenovirus (FAdV-4) in chicken flocks has become endemic in China, which greatly threatens the sustainable development of poultry industry. The development of recombinant FAdV-4 expressing foreign genes is an efficient strategy for controlling both FAdV-4 and other important poultry pathogens. Previous reverse genetic technique for generating the recombinant fowl adenovirus is generally inefficient. In this study, a recombinant FAdV-4 expressing enhanced green fluorescence protein (EGFP), FA4-EGFP, was used as a template virus and directly edited fiber-2 gene to develop an efficient double-fluorescence approach to generate recombinant FAdV-4 through CRISPR/Cas9 and Cre-Loxp system. Moreover, using this strategy, a recombinant virus FAdV4-HA(H9) stably expressing the HA gene of H9N2 influenza virus was generated. Chicken infection study revealed that the recombinant virus FAdV4-HA(H9) was attenuated, and could induce haemagglutination inhibition (HI) titer against H9N2 influenza virus at early time points and inhibit the viral replication in oropharynx. All these demonstrate that the novel strategy for constructing recombinant FAdV-4 expressing foreign genes developed here paves the way for rapidly developing attenuated FAdV-4-based recombinant vaccines for fighting the diseases caused by both FAdV-4 and other pathogens.

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