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Viral Production at the GETV Core

The MWRI GETV Core maintains an inventory of stock plasmids and viral vectors that are available for a nominal fee.

Lentiviral vectors are produced in 293FT cells using a four plasmid system and are replication defective. Lentiviral vectors prepared using this system can be used at BSL2. However, investigators are urged to contact their institutional IBC office for specific details and regulations. Certification of replication-defective status by serial transfer and/or P24 Elisa assay is available upon request. Concentrated lentivirus titer is typically 109 virus particles per ml, but this can be construct dependent. Quantity and pricing information is provided on the order form.

Figure 1: The FUGW lentiviral vector backbone features (1) the HIV-1-flap element to increase viral titer, (2) the WRE sequence to increase transgene transcription, and (3) a self-inactivating promoter (SIN). All lentiviral vectors produced by MWRI-GETV are produced on this lentiviral backbone, which was originally described by Lois C, et al. 2002 Science.

Adeno-associated virus 2 (AAV2) is produced in 293 cells and all vectors are purified for distribution. Purity is confirmed by SDS-PAGE and densitometry. A certificate of analysis is available upon request. AAV vectors prepared in this way are suitable for work at biosafety level 1 (BSL1). However, investigators are urged to contact their institutional IBC office for specific details and regulations. AAV2 titer is typically greater than 1011-1012 genome copies per ml, but this can be construct dependent. Other AAV serotypes are available upon request. Please contact the core manager (shengy@upmc.edu). Quantity and pricing information is provided on the order form.

Adenoviral vectors are produced in 293AD cells and purified for distribution. Adenoviral vectors prepared using this system can be used at BSL2. However, investigators are urged to contact their institutional IBC office for specific details and regulations. Adenovirus titer is typically 1011-1013 virus particles per ml, but this can be construct dependent. Quantity and pricing information is provided on the order form.