Temporal Endogenous Gene Expression Profiles in Response to Polymer-Mediated Transfection and Profile Comparison to Lipid-Mediated Transfection

The current understanding of the molecular factors underlying polyethylenimine(PEI)-mediated transfection is largely unknown. Cationic PEI was used to transfer GFP transgene to HEK293T cells. FACS separation of transfected (GFP positive), untransfected (GFP negative), and untreated cells enabled gene expression profiles to be obtained using Affymetrix® HG-U133A 2.0 microarrays for each cell population. Gene profiles were differentially compared for each population combination. Control (untreated), transfected (GFP positive), and untransfected (GFP negative) RNA samples were purified from HEK293T cells after FACS isolation. Treated experiments were repeated on triplicate days to run 3 microarrays for each cell population. Controls were run on duplicate to give 2 microarrays for each population.