Vector Core

Services

Typically, the Core provides a viral shuttle plasmid to the investigator who inserts the cDNA of interest. Recombinant viral stocks generated from viral plaques are provided to the investigator to evaluate gene expression using the appropriate methodology (either structural or functional assay). Retroviral vectors and packaging cell lines are also available from the Core. Transient transfection using a three-plasmid system is used to produce high titer retroviral supernatants from 293T cells that can be pseudotyped if desired. The Core also supports the use of non-viral vectors for gene transfer by maintaining a repository. A series of plasmid-based vectors containing marker genes, cytokines or polylinkers that were originally developed under the National Gene Vector Laboratory program are available to all investigators following inquiry with the Director.

Please contact the Director for scientific advice and availability of services, pricing, details and scheduling of services.

List of services

Adenovirus services

  • Co-transfection of adenoviral shuttle vectors with adenovirus helper DNA into producer cell lines
  • Plaque purification of viral clones
  • Growth and purification of high titer recombinant stocks
  • Assay of recombinant and wild type virus titers by plaque assay or TaqMan PCR

Viral vectors

Adenovirus

  • E1-replacement; E1, E3 deleted; E1,E3,E4 deleted; E1,E3-deleted ts125 second-generation vectors
  • Targeted Ad vectors (fiber modified)
  • Tet-ON adenovirus vectors (single vector, E1/E3 replacement)
  • Stock vectors (marker viruses, empty vectors) or custom constructs

Viral services

  • Retrovirus (by transient transfection)
  • Lentivirus (by transient transfection)
  • Construction
  • Amplification/propagation
  • Purification
  • Titration (optical density, plaque assay, TaqMan qPCR)
  • Quality control (plaque assay, identity testing, functional testing, RCA testing)

Non-viral vectors

  • Stock vectors
  • Construction of new vectors
  • Propagation (0.05-10L volumes)
  • Purification (Qiagen, CsCl banding or column chromatography)
  • Quality control (testing for endotoxin, protein, RNA and E. coli DNA contamination)
  • Provide viral cloning plasmids for adenovirus, lentivirus and retrovirus vectors
  • GLP manufacturing capability