Gene Transfer and Somatic Cell Engineering Core

Sloan-Kettering Inst Can Research, New York NY

Investigators

Linked publications & trials

Abstract

The Gene Transfer Core (GTC) supports gene transfer studies in Projects 1,2 3 and 4. These projects are[unreadable] highly dependent on achieving efficient gene transfer in primary cells, including hematopoietic progenitor[unreadable] cells, T lymphocytes and dendritic cells. In the upcoming grant cycle, the GTC will mainly focus on the[unreadable] development and optimization of clinical processes and on the implementation of the clinical trials.[unreadable] The specific aims of the Gene Transfer Core are to carry out and/or coordinate: 1. Generation and[unreadable] characterization of high-titer producer cell clones, master cell banks (MCB) for clinical studies 2. Production[unreadable] and titration of 5 to 15 liter batches of clinical viral stocks in semi-closed systems 3. Production and biosafety[unreadable] testing of clinical grade plasmid DNA vaccine for immunization 4. Expansion and transduction of patient cells[unreadable] in semi-closed systems in collaboration with the investigators of each clinical trial 5. Detection of replicationcompetent[unreadable] retrovirus and other biosafety testing in cultured packaging cell clones (MCB), viral stocks and[unreadable] clinical specimen 6. Generation and biosafety testing of clinical AAPCs 7. Detection of oncoretroviral vector[unreadable] integration sites by LM-PCR in patient cells 8. Cell banking, storage of viral stocks, plasmid DNA vaccine[unreadable] and clinical specimens. In addition, the GTC provides essential advisory and training functions for the[unreadable] generation of research grade reagents within the program. The Projects are thus advised or trained on 1.[unreadable] How to construct recombinant oncoretroviral- and lentiviral- vectors, plasmid DNA vectors, siRNA encoding[unreadable] retroviral vectors 2. What packaging cell lines to use 3. How to transfect vector DNA in packaging cells and[unreadable] select producer cell lines 4. What tests to perform to analyze gene expression 5. How to titrate cell-free[unreadable] retroviral stocks by flow cytometry, Southern blot or real time PCR analysis.[unreadable] The GTC is also a repository of numerous reagents and protocols that are freely distributed to all members[unreadable] of the Program who request them. A list of these reagents and protocols will be posted on the newly created[unreadable] Program web portal to facilitate distribution and interactions between the Projects. Vector production,[unreadable] plasmid DNA manufacturing and cell transduction in the GTC decreases the cost of clinical development,[unreadable] ensures high quality and consistency of molecular and cellular processes, and their availability to all[unreadable] investigators in the Program.[unreadable]