Adenovirus/Retrovirus – Services and Fees
Adenovirus
The adenoviral vector used at the VCF is a serotype 5 first generation vector deleted for E1 and E3. The gene of interest is introduced into the vector using a shuttle vector that carries a CMV expression cassette with a LoxP site. Recombination between the vector backbone and the shuttle plasmid is driven by the CRE recombinase, resulting in a high frequency of the appropriate recombinants.
The VCF carries a variety of stock adenoviruses for distribution to registered researchers, and offers a variety of additional services involving adenoviruses:
| Stock | |
|---|---|
| 1 vial of 1x10e11 adenoviral particles | $42 |
| 5 vials of 1x10e11 adenoviral particles | $185 |
| 10 vials of 1x10e11 adenoviral particles | $320 |
| 25 vials of 1x10e11 adenoviral particles | $575 |
| Medium production: 10x15cm plate production, typically yielding 1x10e13 viral particles | $925 |
| Large production: One cell factory, yielding from 1 to 4x10e13 viral particles | $1196 |
| Expansions | |
| Medium scale expansion of your adenovirus or lysate. This is a 10x15cm plate production, typically yielding 1x10e13 viral particles. | $925 |
| Large scale expansion of your adenovirus or lysate. This is one cell factory, typically yielding from 1 to 4x10e13 viral particles. | $1196 |
| Cloning Services & Productions | |
| Adenoviral cloning, when you supply your gene of interest ready for direct insertion into our shuttle plasmid | $626 |
| Adenoviral cloning, when you supply your gene of interest not ready for direct insertion into our shuttle plasmid | $876 |
| Adenoviral cloning, when you do not have the gene of interest, and for siRNA sequence cloning | Call for price |
| Medium scale production of recombinant adenovirus from shuttle plasmid. This is a 10x15cm plate production, typically yielding 1x10e13 viral particles. We cannot guarantee the amounts produced or that the resulting product will be non-toxic. We will make two attempts, and if you wish that we continue after that, the work will be charged regardless of the outcome. | $1860 |
| Large scale production of recombinant adenovirus from shuttle plasmid. This is one cell factory, typically yielding from 1 to 4x10e13 viral particles. We cannot guarantee the amounts produced or that the resulting product will be non-toxic. We will make two attempts, and if you wish that we continue after that, the work will be charged regardless of the outcome. | $2130 |
| Optional Add-on Quality Control | |
| Replication Competent Adenovirus (RCA) testing, available for medium or large expansions or productions | $190 |
| Vector sequencing of vector to ensure presence of insert (first and last 400 bp of insert), available for medium or large expansions or productions | $80 |
| Sterility testing of adenoviral vector, available for medium or large expansions or productions | $100 |
Retrovirus
MFG, the retroviral vector, provides high-level expression of a variety of inserted genes, like cytokines, receptors, enzymes, and growth factors, following non-selective infection of cells. Further modification to enable introduction of multiple genes into the MFG vector was achieved by cloning an internal ribosome entry site (IRES) sequence from encephalomyocarditis virus (EMCV) that allows for translation of a downstream gene in a polycistronic message. The MFG-IRES vectors were shown to efficiently express two genes from the same message using two different secreted marker proteins, IL1Ra (IRAP) and growth hormone. The MFG-IRES vectors have now been used to express three genes such as both subunits of IL-12 and a neomycin resistance gene (neoR) marker gene using two IRES elements.
The VCF provides stock vials of untitered retroviral supernatant to registered researchers, as well as other services involving retroviruses:
| Stock | |
|---|---|
| Small production (2x2 ml vials, untitered) | $223 |
| Medium production (4x2 ml vials, untitered) | $396 |
| Large production (8x2 ml vials, untitered) | $595 |
| Cloning Services & Productions | |
| Retroviral cloning, when you supply your gene of interest ready for direct insertion into our pro-viral plasmid | $626 |
| Retroviral cloning, when you supply your gene of interest not ready for direct insertion into our pro-viral plasmid | $876 |
| Retroviral cloning, when you do not have the gene of interest, and for siRNA sequence cloning | Call for price |
| If you want to add a resistant gene when you do not have the gene of interest | $300 |
| Small production (2x2 ml vials, untitered) | $223 |
| Medium production (4x2 ml vials, untitered) | $396 |
| Large production (8x2 ml vials, untitered) | $595 |
Concerning retroviral orders
- When you order cloning services, you also need to select a small, medium, or large production of the resulting virus.
- We can also supply the pro-viral plasmid so that you may perform the cloning if you wish, and subsequently provide the core with the appropriate amount for production.
- The standard package line is an amphotropic cell line retrovirus that is able to transfect murine and human cells. Some cell types (T-cell, CD34) may require a different envelope. Please contact the VCF to discuss envelope options if using other cell types.
