Characterisation of chromatin states in erythroid cells from a patient with a unique severe HbH genotype (NG Capture C)

Analyses of mutations within the a-globin cluster which downregulate a-globin expression and cause a-thalassaemia provide the basis for genetic counselling and pre-natal diagnosis of this common form of anemia. Understanding the mechanisms by which such mutations cause a-thalassemia has established many of the principles by which mammalian genes are regulated and how this goes awry in human genetic disease. ATAC-Seq and NG Capture-C data from an individual (NSE) with a unique a-globin genotype involving a deletion of the main alpha-globin enhancer on one allele and the --/SEA mutation on the other help to address how the human a-globin cluster is normally regulated. Overall design: To identify which regulatory elements were interacting with the alpha-globin promoters on the functional allele we performed high resolution chromatin conformation capture (3C) using NG CaptureC (Davies 2016). NG CaptureC was performed on primary human cells in triplicate on Day 13 of erythroid differentiation from CD34+ cells isolated from peripheral blood. Capture was performed with biotinylated oligonucleotides targeting sequences adjacent to DpnII sites.