Towards construction of viral vectors based on avian coronavirus infectious bronchitis virus for gene delivery and vaccine development

• Published in: Journal of virological methods Vol. 160; no. 1; pp. 48 - 56
• Main Authors: Shen, Hongyuan, Fang, Shou Guo, Chen, Bo, Chen, Guang, Tay, Felicia P.L., Liu, Ding Xiang
• Format: Journal Article
• Language: English
• Published: Kidlington Elsevier B.V 01.09.2009; Elsevier
• Subjects: Animals; Avian coronavirus infectious bronchitis virus; Biological and medical sciences; Chick Embryo; Chickens; Coronavirus; Dengue Virus - genetics; Dengue virus type 1; Fundamental and applied biological sciences. Psychology; Gene delivery; Genes, Reporter; Genetic Therapy; Genetic Vectors; Genomic Instability; Green Fluorescent Proteins - genetics; Green Fluorescent Proteins - metabolism; Infectious bronchitis virus; Infectious bronchitis virus - genetics; Luciferases, Firefly - genetics; Luciferases, Firefly - metabolism; Microbiology; Reverse genetics; SARS coronavirus; Sequence Deletion; Severe acute respiratory syndrome-related coronavirus - genetics; Techniques used in virology; Viral Proteins - genetics; Viral Vaccines - genetics; Viral Vaccines - immunology; Viral vector; Virology; Gene delivery; Coronavirus; Avian coronavirus infectious bronchitis virus; Viral vector; Reverse genetics; Microbiology; Vaccine; Avian infectious bronchitis virus; Method; Virology; Infection; Virus; Avian infectious bronchitis; Gene; Viral disease; Avian coronavirus infectious bronchitis; Genetics; Coronaviridae; Veterinary; Vector; Nidovirales
• ISSN: 0166-0934; 1879-0984; 1879-0984
• DOI: 10.1016/j.jviromet.2009.04.023

Manipulation of the coronavirus genome to accommodate and express foreign genes is an attractive approach for gene delivery and vaccine development. By using an infectious cloning system developed recently for the avian coronavirus infectious bronchitis virus (IBV), the enhanced green fluorescent protein (EGFP) gene, the firefly luciferase gene and several host and viral genes (eIF3f, SARS ORF6, Dengue virus 1 core protein gene) were inserted into various positions of the IBV genome, and the effects on gene expression, virus recovery, and stability in cell culture were studied. Selected viruses were also inoculated into chicken embryos for studies of foreign gene expression at different tissue level. The results demonstrated the stability of recombinant viruses depends on the intrinsic properties of the foreign gene itself as well as the position at which the foreign genes were inserted. For unstable viruses, the loss of expression of the inserted genes was found to result from a large deletion of the inserted gene and even IBV backbone sequences. This represents a promising system for development of coronavirus-based gene delivery vectors and vaccines against coronavirus and other viral infections in chicken.