Viral Vector Production Core Laboratory
Service and Expertise
In vivo and in vitro viral vector-mediated gene expression
The Core produces popular viral vectors for nonhuman research purpose. The Core has experiences with adenoviral vectors (Ad), adeno-associated viral vectors (AAV), lentiviral vector (LV) and retroviral vectors (RV). However, we currently produce first generation Ad, helper-dependent Ad (HDAd), AAV and LV. We also amplify Ad vectors from viral stocks. The Core provides basic shuttle vectors and expression cassettes. This includes shRNA shuttle vectors for Ad and LV. The customers are required to submit their shuttle vectors ready for transfection or viral stock. In unusual circumstances, the Core assists investigators in cloning into shuttle vectors. Ad vectors are purified over discontinuous CsCl ultracentrifugation. AAV vectors are purified over iodixanol density gradient and are concentrated. LV vectors are concentrated by spin column or by ultracentrifugation. There is no single vector fit for all purposes. The Core personnel are familiar with various viral vector systems and is able to advise the system to fit your need. In addition, we assist investigators to obtain approval for the use of viral vectors in vitro and in vivo.
Expression of fluorescence protein using viral vectors. (A) Mouse pancreatic islets were isolated and infected with helper-dependent adenoviral vector expressing enhanced green fluorescence (GFP) driven by mouse insulin promoter. The image was taken 7 days after infection (magnification x100). (B) Mouse primary hippocampal neurons were infected with adeno-associated virus (serotype 1) expressing GFP driven by elongation factor-1 (EF-1) promoter. The image was taken 5 days after infection (200x). (C) Human hepatoma cell line Hep3B was infected with lentiviral vector expressing dsRed in a 12-well plate, incubated in the presence of puromycin and then expanded into a 6-well plate. The image was taken 8 days after infection (100x). Upper panel: bright field; lower panel: fluorescence image with GFP or Rhodamine filter. (D) Mouse brain. Adeno-associated virus (serotype 6) expressing Green-1 under EF-1 promoter was stereotaxically injected into adult mouse brain (dentate gyrus area). The fluorescence image was taken 7 days after injection. Upper panel: DAPI staining; lower panel: DAPI staining was merged with fluorescence image. Courtesy of Dr. Cecilia Ljungberg.
Director: Kazuhiro Oka, Ph.D.,
Associate Professor, Department of Molecular & Cellular Biology
Baylor College of Medicine
One Baylor Plaza, Alkek N-520.07
Houston, TX 77030