Open Access Thesis
Irwin Martin, Ph.D.
Timothy Cornell, M.D.
Thomas Shanley, M.D.
Association between histones and DNA is crucial for many cellular functions such as gene transcription and epigenetic silencing. Changes to chromatin structure influence gene expression via histone modifications. Chromatin immunoprecipitation (ChIP) is an experimental technique used to investigate the interactions between histones and DNA. ChIP determines the specific location in the genome that histone modifications are associated with, indicating the target of the histone modifiers. Despite the appeal of ChIP as an in vitro technique, there are limitations to its use. There is variability from one preparation to the next, given the many steps and reagents used throughout the technique. No one has been able to demonstrate consistent results in a clinical population using white blood cells. It was established, for the first time, the normal variability of ChIP results in THP-1 cells, Jurkat cells, TF-1a, and human male circulating leukocytes over time for histone modifications H3k4me3 and H3k9Ac.
McDonough, Kelli, "Reproducibility and reliability of chromatin immunoprecipitation in clinical research" (2016). Master's Theses and Doctoral Dissertations. 682.