Eukaryotic Transcriptional and Post-Transcriptional Gene Expression Regulation

Jia Zhang, Klaus H. Kaestner

Narendra Wajapeyee, Romi Gupta

Epigenetic mechanisms have been proposed to contribute to multiple complex diseases, including diabetes mellitus. Dissecting the epigenomic landscape of normal and disease states has greatly expanded our understanding of the regulation of key players in disease pathogenesis and is thus opening doors to novel therapeutic avenues. The human pancreatic islet is a pivotal micro-organ that maintains global glucose homeostasis. The heterogeneity of the islet impedes meaningful in-depth epigenetic analysis using whole islet tissue, because important marks in one cell type are masked by signals from other cell types. We describe here a detailed protocol for the isolation of highly purified endocrine cell subtypes (beta, alpha, and delta cells) from human cadaveric islets, to perform cell-type-specific epigenomic analysis of histone modifications as well as global gene expression profiling.