Construction of stable packaging cell lines for clinical lentiviral vector production

• DOI: 10.1038/srep09021
• Publication Date: 2015-03-12T10:04:17Z
• Authors: Khaled S. Sanber; Sean B. Knight; Sam L. Stephen; Ranbir Bailey; David Escors; Jeremy Minshull; Giorgia Santilli; Adrian J. Thrasher; Mary K. Collins; Yasuhiro Takeuchi
• Keywords: Insert (composites); Transduction (biophysics); Expression cassette

Abstract Lentiviral vectors are useful experimental tools for stable gene delivery and have been used to treat human inherited genetic disorders hematologic malignancies with promising results. Because some of the lentiviral vector components cytotoxic, transient plasmid transfection has produce large batches needed clinical trials. However, this method is costly, poorly reproducible hard scale up. Here we describe a general construction packaging cell lines that continuously vectors. This uses Cre recombinase-mediated cassette exchange insert codon-optimised HIV-1 Gag-Pol expression construct in expressed locus 293FT cells. Subsequently Rev, envelope genome cassettes serially transfected. Vector titers excess 10 6 transducing units/ml can be harvested from final producer clones, which increased 8 TU/ml by concentration. will use all basic investigators who wish