Subconfluent primary human foreskin fibroblast cells are infected with rAAV targeting vectors when is being actively transcribed or replicated in proliferative cells

Abstract

Background reporter gene expression is silenced through density arrest and serum withdrawal. Reporter gene expression is induced by serum stimulation. Single EGFP-Luciferase positive cells are selected using FACS. Cloned cells are expanded and screened for gene targeting events by PCR, Southern blot, and sequencing. FACS histograms are shown for wildtype and rAAV targeted cells following cell cycle entry. FL1-A (EGFP) was plotted against FL2-A (no fluorofor) to center the parent population and allow for selection of the dim EGFP-Luciferase positive cells. The P2 region of the plot indicates the gating used for selection of the EGFP-Luciferase positive portion of the parent population.<p><b>Copyright information:</b></p><p>Taken from "Development of human gene reporter cell lines using rAAV mediated homologous recombination"</p><p></p><p>Biological Procedures Online 2007;9():84-90.</p><p>Published online Jan 2007</p><p>PMCID:PMC2374725.</p><p>Article © by the author(s). This paper is Open Access and is published in Biological Procedures Online under license from the author(s). Copying, printing, redistribution and storage permitted. Journal © 1997-2007 Biological Procedures Online.</p