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Vector Core (Penn)

Director: Johnston, Julie, Ph.D.

Summary:

The Penn Vector Core is a full service viral vector core facility located on the University of Pennsylvania campus. With over a decade of experience in the production of viral-based vectors, the Core has become an important technological resource for investigators, both within and external to Penn, interested in the use of viral based vectors for gene transfer. The main objective of the Core is to provide investigators access to state-of-the-art vector technology for preclinical studies and other basic research applications. Such studies, utilizing carefully designed viral vectors, can provide information critical to the understanding of gene function and development of therapeutic vectors.

The Penn Vector Core specializes in the provision of novel AAV serotype vectors and has the greatest experience in producing novel serotype vectors developed at Penn. AAV1, 7, 8, 9 and rh10 were originally isolated at Penn in the laboratory of Dr. James M. Wilson and first made available to investigators through the Penn Vector Core. Due to its close proximity to the Wilson laboratory, the Penn Vector Core is able to rapidly assimilate new vector technologies and make them available to its users. The Core offers a variety of novel serotype AAV vectors and additional vectors currently under development will be distributed through the Penn Vector. All of the vectors generated by the Penn Vector Core are distributed under material transfer agreement (MTA) to academic, government and non-profit institutions. Corporate users may access novel AAV vector technologies through the Penn co-founded company, REGENXBIO Inc.

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Protocols

  • Quality control standard operating procedures for AAV vectors, Ad vectors, and Lentiviral vectors ( Protocol )

    AAV vectors quality control assays: plasmid characterization, genome copy number titration, infectious titer determination (Taqman TCID50), purity (SDS-PAGE), endotoxin assay (Limulus Amebocyte Lysate)

    Ad vectors quality control assays: physical particle number determination, infectious titer determination (Taqman TCID50), replication-competant adenovirus (RCA) assay, genome structure analysis, endotoxin assay (Limulus Amebocyte Lysate)

    Lentiviral/retroviral vectors quality control assays: infectious DNA titer assay, replication-competent lentivirus (RCL), endotoxin assay (Limulus Amebocyte Lysate)

Services

  • Adeno-associated viral vector services ( Material production service )

    "A number of novel AAV vectors were originally isolated and developed at Penn in the laboratory of Dr. James Wilson and first made available to investigators through the Penn Vector Core. Due to its close proximity to the Wilson laboratory, the Penn Vector Core is able to rapidly assimilate new vector technologies and make them available to its users. The Core offers a variety of novel serotype AAV vectors and additional vectors currently under development will be distributed through the Penn Vector.

    The Core currently offers 8 AAV vector serotypes, with additional serotypes also available. The tropism of selected AAV serotypes is shown in the following table with an X shown where efficacy has been demonstrated. For targets not listed, it may be necessary to conduct pilot serotype selection studies to determine the serotype of choice. We recommend serotype selection studies be performed in vivo since the in vitro transduction efficiencies of many of the serotypes below does not accurately reflect in vivo performance. For investigators wishing to discuss the serotype of choice for their application, please contact the Director of the Penn Vector Core.

    AAV1: skeletal muscle, heart, lung, CNS/eye
    AAV2: CNS/eye
    AAV5: lung, CNS/eye
    AAV6: skeletal muscle, heart, lung, CNS/eye, liver, adipose
    AAV7: skeletal muscle, CNS/eye, liver
    AAV8: skeletal muscle, CNS/eye, liver, adipose
    AAV9: skeletal muscle, heart, lung, CNS/eye, liver, adipose
    AAVrh10: skeletal muscle, lung, liver

    The Penn Vector Core is pleased to offer four scales of AAV production services."

  • Adenoviral vector services ( Material production service )

    "Recombinant adenoviral vectors are one of the most efficient viral vectors for gene delivery, both in vivo and in vitro, due to their high transduction efficiency, broad host range, ability to infect non-dividing cells, and potential for generating high titer virus. Applications for this vector include gene therapy, particularly in cases where high level, short term gene expression is required, vaccine applications and basic gene transfer studies. Recombinant adenoviral vectors are produced by direct cloning, produced in complementing cell lines and purified by cesium chloride gradient ultracentrifugation."

  • Lentiviral vector services ( Material production service )

    "The Penn Vector Core offers a single scale* of lentiviral vector production."

  • Vector support services ( Material processing service )

    "The rates for support services including cloning (one-step), DNA amplification (for routine scale production) and DNA structure and sequence analysis are provided in the table below. Note that DNA structure and sequence analysis services are required if in support of custom cloning services provided by the core and include full genome (eg. from AAV 5’ ITR to 3’ ITR) sequence analysis as well as thorough restriction enzyme analysis."


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Last updated: 2016-04-22T12:43:39.420-04:00

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