The Vector Core will provide state-of-the-art vector services for the pre-clinical efforts of this program. Vectors for clinical studies will not be provided. To meet these goals, the Vector Core is organized so that each staff member has well-defined tasks while cross-training is provided. (1) The Vector Core researchers will produce, purify and quality control replication-defective chimpanzee (chimp) adenovirus (Ad) vectors and Ad vectors of the human serotype 5 (AdHu5). Specifically they will conduct all of the steps needed to generate new recombinant Ad vectors, which involves cloning of novel inserts (sequence to be provided by the investigators) into shuttle vectors and from there into the molecular viral clones. Replication-defective Ad virus will then be rescued and expanded on HEK 293 cells. Upon purification, virus is titrated both for content of virus particles and infectious units. The Ad vectors in addition will undergo a number of vigorous quality controls including testing for genetic integrity, contamination with replication-competent Ads or LPS. Furthermore upon request vectors will be tested for transgene product expression and for genetic stability. (2) The Core will expand, purify and quality control Ad vectors from already available virus stocks. (3) The Core will produce live virus for the projects. (4) The Core will be responsible for storage of vectors and for maintaining adequate inventories and databases. The database will list all of the vectors that have been produced, including results of titration and quality control assays. The database will list the status of newly requested vectors and it will list the availability of vectors. The database will be updated weekly and shared electronically with the Project Leaders. (5) The Core will assist Project 1 in transfer of technology to subcontractors involved in GMP vector production and release testing.
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