Establishment of a Simple and Efficient Reverse Genetics System for Canine Adenoviruses Using Bacterial Artificial Chromosomes
Open Access
- 15 July 2020
- Vol. 12 (7), 767
- https://doi.org/10.3390/v12070767
Abstract
Canine adenoviruses (CAdVs) are divided into pathotypes CAdV1 and CAdV2, which cause infectious hepatitis and laryngotracheitis in canid animals, respectively. They can be the backbones of viral vectors that could be applied in recombinant vaccines or for gene transfer in dogs and in serologically naïve humans. Although conventional plasmid-based reverse genetics systems can be used to construct CAdV vectors, their large genome size creates technical difficulties in gene cloning and manipulation. In this study, we established an improved reverse genetics system for CAdVs using bacterial artificial chromosomes (BACs), in which genetic modifications can be efficiently and simply made through BAC recombineering. To validate the utility of this system, we used it to generate CAdV2 with the early region 1 gene deleted. This mutant was robustly generated and attenuated in cell culture. The results suggest that our established BAC-based reverse genetics system for CAdVs would be a useful and powerful tool for basic and advanced practical studies with these viruses.Funding Information
- Japan Society for the Promotion of Science (Scientific Research (A) 18H03971)
This publication has 47 references indexed in Scilit:
- A Novel Chimpanzee Adenovirus Vector with Low Human Seroprevalence: Improved Systems for Vector Derivation and Comparative ImmunogenicityPLOS ONE, 2012
- An Update on Canine Adenovirus Type 2 and Its VectorsViruses, 2010
- Generation of E3-deleted canine adenovirus type 2 expressing the Gc glycoprotein of Seoul virus by gene insertion or deletion of related terminal region sequencesJournal of General Virology, 2010
- Adenovirus E1B 55-Kilodalton Protein: Multiple Roles in Viral Infection and Cell TransformationJournal of Virology, 2009
- Infectious canine hepatitis: An “old” disease reemerging in ItalyResearch in Veterinary Science, 2007
- Development of nonhuman adenoviruses as vaccine vectorsVaccine, 2006
- Telomerase-dependent oncolytic adenovirus for cancer treatmentGene Therapy, 2003
- A mutant oncolytic adenovirus targeting the Rb pathway produces anti-glioma effect in vivoOncogene, 2000
- Efficient Construction of a Recombinant Adenovirus Vector by an ImprovedIn VitroLigation MethodHuman Gene Therapy, 1998
- Emergence of Early Region 1-Containing Replication-Competent Adenovirus in Stocks of Replication-Defective Adenovirus Recombinants (ΔE1 + ΔE3) During Multiple Passages in 293 CellsHuman Gene Therapy, 1994