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Adenoviral Vector

Adenoviral vectors were developed from human sera type V adenovirus, which are replication defective by deleting a E1 and E3 regions. The gene of interest is usually inserted in E1 region of adeno-genome. Adenoviral vector are very efficient to infect non-dividing and dividing cells for transient gene expression.

Adenoviral vector for gene overexpression

The VVC has AdenoEasy system available for our clients. This system requires to subclone the genes of interest into the shuttle vector, in which a CMV promoter is used to drive the gene expression. This shuttle vector containing the gene of interest will be incorporated into the adeno-genome through the homologous recombination in a adeno-genome transformed bacteria strain.

There is a doxcycline inducible adenoviral vector system available from VVC. This is a second generation based inducible system and developed by VVC recently. The gene of interest will be subcloned into a shuttle vector containing a tetracycline regulated Tet-tight promoter. This shuttle vector will be incorporated into the E1 region of adeno-genome. We have a reverse transactivator rtTA-M2 in a separated shuttle vector. The rtTA-M2 is incorporated into E3 region of adeno-genome. The VVC use a phage packaging system to obtain the recombinant adenogenome with a 100% recombination rate. The VVC has tested this system successfully using EGFP reporter gene (Fig.1). Fig. 1: Second generation doxcycline inducible adenoviral vector

Adenoviral shRNA mediated adenoviral vector knockdown

The VVC has an AdenoEasy based shRNA vector, which can be used for gene knockdown studies. The hairpin structure can be inserted HindIII and BamHI site and driven by polymerase III promoter H1.

Detection of replication competent adenoviral vector

The VVC has the IBC approved method for detecting replication competent virus by infection non-permissive cell lines and PCR based method.

Services

The VVC has the following services available for UTHSC faculty members:

  1. Design lentiviral and adenoviral vectors for overexpression and knockdown or inducible expression and knockdown
  2. Produce lentiviral and adenoviral vectors
  3. Concentrate and purify lentiviral and adenoviral vectors
  4. Training on viral vector manipulation
  5. Detect replication competent virus
  6. Distribute recombinant reporter virus


Service Requisition Form and Price Schedule

Personnel

Viral Vector Core Laboratory, Scientific Director
Dr. Lorraine Albritton, Ph.D, Professor
Department of Microbiology, Immunology & Biochemistry


Viral Vector Core Laboratory, Director/manager
Junming Yue Ph.D. Assistant Professor
Department of Physiology

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