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Helper-dependent adenovirus (HDAd), also known as Gutless adenovirus (GLAd), has been considered as the last-generation adenovirus. HDAd is constructed by deletion of all viral genes, resulting in no expression of adenoviral proteins. The only remained adenoviral sequences include ITRs, the adenoviral packaging signal, and a small untranscribed portion of the E4 region.
HDAd has many notable characteristics as a gene delivery vector. HDAd still retains the advantages of early generations of adenovirus such as a broad cell/tissue tropism, high infectivity, high transgene expression, and an absence of integration into the host genome. Since it is constructed following the deletion of all the adenoviral genes, HDAd exhibits a very large accommodation capacity for transgenes, which is approximately 30 kb, making it possible to deliver large genes as well as multiple genes. The absence of all adenoviral genes also enables HDAd to mediate long-term transgene expression in host organisms with minimal immunogenicity.
Figure 1. Production of HDAd using the Cre/loxP system (Robin JP, et al., 1996)
Since HDAd is devoid of all genes from the adenovirus genome, the production of HDAd is dependent upon a helper adenovirus that provides all viral proteins for virus packaging. Currently, the most efficient and widely used strategy for producing HDAd is based upon the Cre/loxP recombination system. Briefly, the HDAd vector is transfected into 293 cells expressing the Cre recombinase. After transfection the helper virus is added for providing all the needed viral proteins during virus packaging.
Creative Biogene offers custom services for producing Helper-dependent adenovirus (HDAd). Our HDAd production is based on the Cre/loxP recombination system. Because the packaging signal of the helper adenovirus is floxed and excised by Cre recombinase, only HDAd vector genome can be packaged into adenoviral particles. The produced HDAds are purified by density gradient centrifugation and can be used for downstream animal injections.
-Infect dividing and non-dividing cells
-Transient high-level protein expression
-Express multiple genes in a single virus
-Very large capacity, up to ~30kb
-High virus titer can be produced
-No genome integration
-Lower host immune response due to no viral genes
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