Viral Vectors

Constructing and packaging viral vectors for experimental gene delivery Read More

Notice re Hope Center Subsidy

Beginning with the August 2013 billing cycle, the Hope Center subsidy will apply only if the billing PI is a Hope Center faculty member.

About

The Hope Center Viral Vectors Core provides lentiviral and AAV vectors and other services to Washington University investigators on a fee-for-service basis. Investigators from nonprofit/academic institutions outside Washington University may use Core Services as well. The Core has special expertise in the use of viral vectors for neuroscience applications. The Core also supervises shared molecular and cell culture equipment in the Hope Center. The goals of the Core are to provide the highest quality viral vectors available, to serve as a repository for the viral resources and expertise available at Washington University and to develop new vectors and reporters for neuroscience research.

Scroll down to learn about services and vectors available, as well as suggested language for acknowledging core use.

Services

The Core will assist investigators with the design and construction of lentiviral vectors, and with small or large scale viral packaging. We can provide viruses expressing marker proteins for use in testing viral expression in different cell types and organs. Basic reporter vectors are available for research purposes to Washington University investigators; some transgenes and vectors may require that the investigator obtain permission from the source lab. Investigators should obtain Recombinant DNA approval from Environmental Health and Safety prior to using these vectors in their laboratories. Labs that have not yet obtained approval can perform some experiments in the Core facility with training and supervision by Core staff. Vectors are not available for for-profit or human diagnostic applications.

Lentiviral Vectors

Lentiviral vectors are now widely used for modeling brain diseases, for genetic pharmacology and for delivery of potential therapies into animal models. The Hope Center Viral Vectors Core uses third generation HIV-1 based lentiviral vectors. These vectors have been used successfully in rodent models of lysosomal storage diseases (Hofling et al., 2004) in cultured neurons (Lee et al., 2004) and in PNS cell types such as Schwann cells and dorsal root ganglion cultures (Abdulkadir et al., 2002; Modur et al., 2002; Araki et al., 2004; Schalch et al., 2004; Le et al., 2005; Bermingham et al., 2006). Third generation lentiviral vectors offer several advantages: they readily transduce CNS neurons and glia, lymphocytes, endothelial cells and stem cells; they have very low toxicity, induce a minimal immune response, and are expressed for prolonged periods (6 months or longer, Blomer et al., 1997). Vectors used currently have a moderate cloning capacity (up to 8 kb) and can be generated with dual promoters or with internal ribosome entry sites (IRES) to enable expression of two transgenes from the same mRNA (e.g., the FCIV vector used here, see Araki et al., 2004).

Lentiviral vectors have also been very useful in the delivery of silencing RNAs. The core can help investigators design and construct new short hairpin RNAs (shRNA) for delivery in lentiviral vectors, or can package lentiviral shRNA vectors obtained from other sources, such as Sigma’s Mission shRNA.

Available lentiviral vectors

Adeno-associated virus (AAV) vectors

Adeno-associated virus is nonpathogenic in humans. Recombinant AAV vectors typically do not elicit a cellular immune response. AAV vectors are able to spread widely in vivo after injection and maintain long-term expression of transgenes. Various serotypes of AAV vectors allow tissue specific delivery of genes of interest (Cearley et al., 2005; Harding et al., 2006). AAV vectors have become increasingly important in neuroscience research and clinical applications.

AAV vector serotypes 1, 2, 5, 6, 7, 8 and 9 are available in the Core.  For AAV vector serotypes 7, 8 and 9, users must obtain an MTA from University of Pennsylvania before use.  The following promoters are available for each serotype of AAV vectors: CAB (chicken β actin promoter with CMV enhancer), SYN (synapsin), and GFAP (glial fibrillary acidic protein).  We will work with users to develop custom promoter constructs if needed.

Acknowledging the Core

The Viral Vectors Core is supported by the grants below as well as the Hope Center for Neurological Disorders. Publications that include data made possible by the core should acknowledge grants as appropriate.  Note that these acknowledgements are crucial in tracking success of the core and obtaining future funding.  Suggested language is included below.

  • NIH Neuroscience Blueprint Interdisciplinary Center Core Grant P30 NS057105

Suggested language to include in acknowledgements: “This work was supported by the Hope Center Viral Vectors Core and a P30 Neuroscience Blueprint Interdisciplinary Center Core award to Washington University (P30 NS057105).”

Feedback

We always strive to improve services and resources offered by the Hope Center Cores.  To provide feedback, see contact information below.

Core Coordinator: Mingjie Li
Faculty Supervisor: Joy Snider

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Contact Information

Initiating Use

Click below to learn more about available vectors, or for a new virus request form.  For questions, contact Mingjie Li.

New virus/service form

Available vectors

Pricing

Charges for Hope Center investigators are subsidized by the Hope Center.  Note that the P30 Blueprint subsidy will not apply to orders placed on February 1, 2013 or later.

Current usage fees