Adenovirus can be used as a vector to express heterologous genes in dividing or non-dividing cells. Adenovirus vectors are deleted for the E1A and E1B genes, and their replication is therefore impaired in most cell types. Additional deletions in the E3 and E4 regions can increase the cargo capacity to about 11 kb foreign DNA. Expression cassettes are inserted most of the time in place of the E1 region, less often in the E3 region, and rarely in the E4 region.
OD260 Inc can construct any type of adenovirus vector according to your needs, as long as it does not require a BL3 facility. We can also help you with the design of your vector.
- Transient gene expression in vitro, especially in cell lines that are refractory to other transfection methods
- Transient gene expression in vivo
- Multi-cistronic vectors: expression of two or more transgenes from the same vector
- Vectors containing two genes of interest under the control of their own promoter
- Vectors containing your gene of interest in the E1 region and a reporter gene in the E3 region or vice-versa
- Vectors containing inducible expression systems
- Vectors expressing a regulator and a regulated gene
- Vectors expressing two genes involved in the same pathway
- Vectors expressing two genes whose products interact with each other
- Vaccine vectors
- Cloning systems: AdenoQuick1.0, AdenoQuick2.0, AdenoZAP, or co-transfection method
- Backbone: Ad5
- Packaging signal: complete (all seven "A" repeats) or partial (repeats A1-A5)
- E1 region: WT or deleted 3.1 kb (bp 354-3503) with multiple cloning site
- E3 region: WT, or deleted (1.6 or 2.7 kb) with multiple cloning site
- E4 region: WT, or deleted (1.2 or 2.8 kb) with multiple cloning site
- Fiber: WT Ad5, hybrid Ad5/3 or Ad5/35
- Maximum cargo capacity: ~ 11.2 kb
- A plasmid DNA containing your transgene, possibly inserted into an expression cassette including a promoter and polyadenylation signal. If necessary, we can synthesize your transgene or even the entire expression cassette
- The sequence of your transgene, or at least a restriction map.
The service includes
- Construction of a shuttle vector containing your expression cassette
- (AdenoQuick™ system only: construction of a cosmid containing the entire sequence of your recombinant virus)
- Virus recovery by transfecting the DNA into helper cells
- Virus clone isolation by plaque assay
- Confirmation of virus identity at the DNA level
You can customize your request: for example, you can decide to construct the shuttle vector yourself and send it to us for virus recovery and characterization.
- The crude virus lysates of three clones of your recombinant virus
- A complete report describing the work performed and demonstrating the identity of the virus we are sending to you
- About 4 weeks, depending on how many cloning steps in E. coli are necessary.
The use of adenovirus as a genetic vector requires adequate containment equipment and practices. Biosafety Level 2 (BL2) is appropriate for most customers; however, for some, use of BL3 or higher level containment facility may be required. O.D.260 Inc. will not carry out projects that require BL3 (or above) containment.