Improved DNA methylation analysis via enrichment of demethylated cells expressing an X-inactivated transgene.
The demethylating drug 5-aza-2'-deoxycytidine (5-aza-2dC) is frequently used to investigate the effect of global DNA demethylation on gene expression in cultured mammalian cells. Here, we describe a method that uses the reactivation of an X-inactivated green fluorescent protein (GFP) transgene as a marker to enrich for cells that have undergone drug-induced demethylation. By combining it with microarray gene expression profiling, we demonstrate the method's utility in identifying genes activated by global DNA demethylation.
Vallender
Tammy W
TW
University of Chicago, Chicago, IL 60637, USA.
Lahn
Bruce T
BT
D000818:Animals; D000831:Animals, Newborn; D001374:Azacitidine / Q000494:pharmacology; D002478:Cells, Cultured; D019175:DNA Methylation; D005260:Female; D005347:Fibroblasts / Q000187:drug effects / Q000378:metabolism; D005434:Flow Cytometry; D020869:Gene Expression Profiling; D005786:Gene Expression Regulation / Q000187:drug effects; D020868:Gene Silencing; D049452:Green Fluorescent Proteins / Q000378:metabolism; D051379:Mice; D020411:Oligonucleotide Array Sequence Analysis; D009924:Organ Culture Techniques; D013997:Time Factors; D019076:Transgenes; D014960:X Chromosome
D049452:Green Fluorescent Proteins; D001374:Azacitidine