Targeted repression of the IE175k promoter by synthetic zinc-finger proteins fused to the CD of Dnmt3a

Copyright information:Taken from "Chimeric DNA methyltransferases target DNA methylation to specific DNA sequences and repress expression of target genes"Nucleic Acids Research 2006;35(1):100-112.Published online 06 Dec 2006PMCID:PMC1761428.© 2006 The Author(s) () The IE175-Luc or IE175mut-Luc reporter plasmids were co-transfected with 6F6, 6F6-3a, B1-3a or 6F6-3a-E74A into HEK293T cells. The IE175mut promoter does not contain the binding site for the engineered zinc finger protein B1/6F6. The Dnmt3a-E74A variant is catalytically inactive. The luciferase activity of reporter gene co-transfected with 6F6 was set to 100%. The error bars indicate the standard deviations of at least three independent experiments. () Competition between 6F6-3a and 6F6. To investigate the targeting function of the zinc-finger DBD, constant amounts of IE175-Luc and 6F6-3a were co-transfected with different concentrations of a construct expressing only 6F6, while the total amount of DNA in each transfection was normalized by adding an empty vector.