Introduction:
Penn Vector Core is an academic core facility located at the University of Pennsylvania (UPenn) that provides viral and non-viral based vectors for preclinical studies. The main objective of the Core is to provide investigators access to state-of-the-art vector technology for gene transfer and other translational research applications. In support of the NHBLI Gene Therapy Resource Program, the Penn Vector Core will provide research grade viral vectors derived from adeno-associated virus (AAV), adenovirus, lentivirus and plasmid-based vectors.
The Core generates both custom vectors designed according to the needs of the investigator as well as ready-made reporter gene vectors. An integral part of the Vector Core is its robust quality control program which is carried out by a separate Quality Control Group. Quality control assays have been developed and optimized for all the vectors produced by the Core and criteria are in place which must be met before the release of any vector produced in our facility. The Vector Core maintains close contact with research laboratories at UPenn and is able to rapidly assimilate new vector technologies.
Service available through this Core Laboratory also includes immunology assays related to the use of preclinical-grade vectors. The work is conducted in a BSL-2 facility under research-grade (non-GLP) conditions which is appropriate for some applications but may not be appropriate for all. Laboratory staff can provide assistance to applying investigators in addressing any questions.
Types of Services:
The Penn Vector Core offers a full range of services and strives to continue to improve, expand, and develop new services that would be of use to investigators interested in vector-mediated gene delivery. The Core offers AAV vectors pseudotyped with a variety of different capsid serotypes, adenoviral vectors containing various backbone deletions, and lentiviral vectors pseudotyped with a range of envelope glycoproteins. In addition to vector production services, the Core provides a number of plasmids and cell lines as well as consultation prior to any request for services. Details for the main services offered by Penn Vector Core are provided below:
Penn Gene Therapy Program Link - http://www.med.upenn.edu/gtp/
Penn Vector Core Laboratory Link - http://www.med.upenn.edu/gtp/vector_core.shtml
Penn Vector Core Contacts Link - http://www.med.upenn.edu/gtp/VectorCoreContact.shtml
- Transduction inhibition based neutralizing antibody assay (AAV, Adenoviral and Lentiviral vectors)
- A sensitive and generic assay developed by the Core and characterized by an easy and quantifiable LacZ read-out in the presence and absence of neutralizing antibodies
- Comparison between different serotypes and cross-reactivity possible
- Can be used with all known AAV and adenoviral vectors and with VSV-G pseudotyped lentiviral vectors
- Interferon gamma (IFNγ) ELISpot to adenoviral vectors, AAV capsid and transgene products (in non-human primate, mouse, rat, ovine, bovine, equine, feline, canine and porcine)
- Intracellular cytokine staining (ICS) to Adv virus, AAV capsid and transgene product (in mice and non-human primates, 5 colors: IFNγ, IL2, TNFα, CD4, and CD8)
- An extensive stock of custom made peptide libraries (15 mers with 10 aminoacid overlap) to different AAV serotypes (AAV1, AAV2, AAV7, AAV8 AAVrh32.33) and to two Adenovirus Hexons (AdV-H5 and SAdV-24 [20 mers with 15 aminoacid overlap])
- Full access to a broad range of novel adenovirus (30 serotypes) and AAV vectors (more than 100) that can be used for T cell detection in both IFNγ and ICS assays
- Synthetic peptide libraries (required for transgene specific T-cell response detection)
- — Library for the commonly used reporter gene EGFP (15 mer with 10 aminoacid overlap)
available for this purpose
- — Investigators may also provide a targeted peptide library
- — Investigators may request the Core order the synthesis of a specific peptide library.
FAQs
For all types of RSAs for laboratory services
- The scientific research in gene therapy that is the subject of the RSA must be consistent with the NHLBI Mission (http://www.nhlbi.nih.gov/about/org/mission.htm) or, if the research addresses a disease area consistent with the mission of another NIH Institute, that Institute must agree to cover the cost of the service.
- The investigator must complete the on-line Registration Form on the GTRP main web page and receive approval by the NHLBI Gene Therapy Group (GTG) allowing the investigator to submit an RSA.
- RSA approval and implementation is contingent upon availability of funds in the GTRP.
- Services provided through an approved RSA are at no cost to the investigator.
- The various types of RSAs are independent of one another. An applicant Investigator may submit any type of RSA for a qualified gene therapy research project regardless of past or future utilization of GTRP resources for the project.
In addition, for Preclinical Vector Production
- The applicant must be working in basic or translational gene therapy research in a disease area funded by the NIH.
- Funding must already be secured, and demonstrated in writing, for the project for which the vector is requested.
A Request for Service Application (RSA) is the mechanism for applying for GTRP resources. Investigators must register on-line and be approved by the NHLBI Gene Therapy Group in order to access the Request for Service Application.
In order for the laboratory to meet the vector production requirements of the study, the applicant investigator will be asked to provide such information as:
- Vector type requested (AAV, adenoviral vector or lentiviral vector)
- Vector serotype or pseudo type required (if known)
- Information about the expression cassette requested (promoter, transgene, polyA)
- Details on any material to be submitted for vector production (plasmid, lysate, purified vector)
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