Vector Development and Production

Core Director:
Miguel Sena-Esteves, Ph.D.
Email: esteves@email.chop.edu
Phone: 617-724-8917

Core functions: This Core will advise investigators on HSV amplicon and/or retrovirus vectors most appropriate for their research needs based on relative infectivity of cells of interest, use for culture or in vivo studies, promoter considerations, hypotheses being addressed, and appropriate reporter genes. The Core will provide appropriate amplicon or retrovirus plasmid backbones and construction information and assistance to investigators, as well as packaging vectors for them.

Investigators using the basic amplicon backbone will be supplied with amplicon plasmids that contain one of the following reporter genes under an immediate early (IE) HSV promoter: enhanced green fluorescent protein (eGFP), red fluorescent protein (dsRFP), luciferase, alkaline phosphatase or lacZ and a multiple cloning site (MCS) downstream of a cytomegalovirus (CMV) IE promoter. Other vector backbones are available for longer-term reporter gene expression, through a CMV IE promoter, in which case the transgene is under the Rous sarcoma virus (RSV) or mammalian promoters.

Investigators will be responsible for inserting their genes of interest into the MCS, validating correct construction by restriction analysis, testing bioactivity by transfection assays (when possible), and supplying clean maxi-prepped DNA for packaging.

The Core will maintain high quality stocks of HSV BAC DNA for packaging and will carry out packaging and “purification”, titering and allocation of amplicon vectors for the investigators.