Impact of exogenous gene insertion at different influenza genome sites
10.3760/cma.j.cn112309-20241231-00454
- VernacularTitle:流感病毒基因组不同位点插入外源基因对病毒特性影响的研究
- Author:
Hao WU
1
;
Chenyan ZHAO
;
Xi WU
;
Li ZHANG
;
Jiayou ZHANG
;
Yongxin YU
;
Weijin HUANG
Author Information
1. 中国食品药品检定研究院艾滋病性病病毒疫苗室，国家药品监督管理局生物制品质量研究与评价重点实验室，北京 102629
- Publication Type:Journal Article
- Keywords:
Influenza virus;
Virus modification;
Gene integration;
Fluorescent labeling;
In vivo imaging
- From:
Chinese Journal of Microbiology and Immunology
2025;45(5):394-399
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To investigate the impact of inserting an exogenous gene, NanoLuc (Nluc), at different sites in the influenza virus genome on viral properties and analyze the expression stability of the exogenous gene both in vitro and in vivo. Methods:Using molecular cloning techniques and reverse genetics, eight recombinant influenza viruses were constructed by inserting the exogenous Nluc gene into the gene segments encoding hemagglutinin (HA), neuraminidase (NA), non-structural protein (NS), and polymerase basic protein 1 (PB1). Viral replication capacity was evaluated by hemagglutination and plaque assays. Nluc expression in infected cells was monitored by fluorescence imaging. The potential impact of the exogenous gene insertion on viral infectivity was examined in a mouse infection model. Independent samples t-test were used for statistical analysis. Results:The recombinant viruses with insertions in the HA, NA, and NS gene segments generated fluorescent signals in the first generation of rescued viruses and demonstrated replication capabilities in plaque and hemagglutination assays. The recombinant viruses based on the NA and NS genes were capable of stably expressing Nluc across different generations, and exhibited correct fluorescent distribution patterns in mouse infection experiments. Meanwhile, the NS gene-based recombinant virus demonstrated superior stability in the mouse model.Conclusions:This study demonstrates that the NS gene segment of influenza virus can serve as an effective insertion site for exogenous genes without impairing the viral replication or infectivity, and the recombinant virus constructed based on it exhibits high integration stability and substantial application potential.
