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Adenovirus and Recombinant Adenovirus

Wild type human adenovirus
Recombinant adenovirus
Advantages using Recombinant Adenovirus


Biology of wild type human adenovirus (type 5)

Wild-type adenoviruses, including human adenovirus type 5, are associated with a number of mild disorders, such as respiration infections in the elderly or children.

The entire adenovirus genome is a linear, double-stranded DNA of approximately 36Kb. There approximately 2 phases of adenovirus transcription, namely early and late phases, which occur before and after replication. The early-transcribed regions are E1, E2, E3 and E4.

The E1 gene products, including E1A and E1B, are involved in the replication of the virus. The E2 proteins provide the machinery for viral DNA replication and transcription of late genes. Most of the E3 proteins are involved in modulating the immune response of infected cells. The E4 gene products are involved in the metabolism of virus messenger RNA and provide functions that promote virus DNA replication and shut-off of host protein synthesis.

The prime receptor for human adenovirus is the Coxsackie/Adenovirus Receptor (CAR). Once inside the cell, the virus translocates to the nuclear pore complex, where the viral DNA is released into the nucleus. Transcription and replication take place in the nucleus of the infected cell.



Recombinant Adenovirus (DE1/E3)

To ensure replication deficiency of the virus, the E1 region is deleted allowing it to safely be used as a gene delivery tool. To accommodate larger recombinant genes (up to 8 Kb), 1st generation adenoviruses are both E1 and E3 deleted (E1/E3), since the E3 region is not essential for in vitro viral growth.

The adenovirus vector is able to deliver genes with 100% efficiency to a wide selection of cell types including dividing or non-dividing cells, or primary cells or cell lines. This ability far surpasses the gene delivery efficiencies of lipid-based transfection approaches or other viral-based gene delivery systems.



Advantages using Recombinant Adenovirus

There are many advantages in using an adenovirus to introduce genetic material into host cells. :

Recombinant adenovirus:

  • Represents a homologous system for human genes: adenoviral vectors use a human virus as vector and human cells as host. Therefore, human proteins have identical post-translational modifications as native proteins.
  • Has the ability to infect most mammalian cell types (both replicative and non-replicative)
  • Accommodates reasonably large transgenes (up to 8 kb)
  • Allows high expression of the recombinant protein
  • May be grown at high titer (1010 VP/mL, which can be concentrated up to 1013 VP/mL)
  • Is well tolerated, with post-infection viability of the host cells being almost 100%
  • Remains epichromosomal, i.e. does not integrate into the host chromosome so does not inactivate genes or activate oncogenes
All these have made recombinant adenovirus the vector of choice for functional genomics research, protein-over-expression and pre-clinical studies.

 

 

Vector BioLabs offers many adenovirus services to fit your research needs, please inquire for details.


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