Rapid, solid-phase based automated analysis of chromatin structure and transcription factor occupancy in living eukaryotic cells.
Transcription factors, chromatin components and chromatin modification activities are involved in many diseases including cancer. However, the means by which alterations in these factors influence the epigenotype of specific cell types is poorly understood. One problem that limits progress is that regulatory regions of eukaryotic genes sometimes extend over large regions of DNA. To improve chromatin structure-function analysis over such large regions, we have developed an automated, relatively simple procedure that uses magnetic beads and a capillary sequencer for ligation-mediated-PCR (LM-PCR). We show that the procedure can be used for the rapid examination of chromatin fine-structure, nucleosome positioning as well as changes in transcription factor binding-site occupancy during cellular differentiation.
Ingram
Richard
R
Molecular Medicine Unit, University of Leeds, St James's University Hospital Leeds LS9 7TF, UK.
Tagoh
Hiromi
H
Riggs
Arthur D
AD
Bonifer
Constanze
C
D000818:Animals; D001665:Binding Sites; D002460:Cell Line; D002843:Chromatin / Q000737:chemistry* / Q000378:metabolism*; D018983:DNA Footprinting; D017931:DNA Primers; D016762:Genes, fos; D051379:Mice; D016133:Polymerase Chain Reaction / Q000379:methods*; D011401:Promoter Regions, Genetic; D017422:Sequence Analysis, DNA; D014157:Transcription Factors / Q000378:metabolism*
D002843:Chromatin; D017931:DNA Primers; D014157:Transcription Factors