University of Pittsburgh Cancer Institute (UPCI)

CCSG Acknowledgement

Required CCSG Acknowledgement

The NCI requires that publications acknowledge the UPCI CCSG support, and they are tracking compliance. If a UPCI CCSG-supported Shared Resource provided data used in your publication, please include the following statement in the acknowledgment section of your publication(s):

"This project used the UPCI [insert name(s) of shared resource(s)] that [is/are] supported in part by award P30CA047904."

Shared Resource Directors: Please make sure to include this statement on all of your order forms, contracts, etc. as a reminder to your users to acknowledge the UPCI CCSG support.


Adenovirus/Retrovirus – Services and Fees

Adenovirus

The adenoviral vector used at the VF 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 VF 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

  1. When you order cloning services, you also need to select a small, medium, or large production of the resulting virus.
  2. 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.
  3. 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 VF to discuss envelope options if using other cell types.