Investigating the Role of Kdm3b in the Acquisition of Pluripotency

Abstract

Induced Pluripotent Stem Cells are somatic cells that have been reprogrammed to acquire the properties of embryonic stem cells by the overexpression of various proteins. These somatic cells have acquired pluripotency by modifying their epigenome, the chemical marks on their DNA and histone tails. These epigenetic modifications are perfonned by various epigenetic enzymes. A family of histone demethylases, the Kdm3 's (Kdm3a, Kdm3b, Kdm3c), are of particular interest to the Sridharan lab, as it has been shown that cells lacking Kdm3b have a significantly reduced reprogramming efficiency, but cells lacking Kdm3a and Kdm3c do not. The goal of my project was to determine the structural aspects of Kdm3b that result in its specific behavior in reprogramming. This was done by creating overexpression vectors that possess Kdm3a, Kdm3b, as well as two mutant Kdm3a/Kdm3b combinations, transducing these overexpression vectors into cells lacking the Kdm3b enzyme, and observing how efficiently these cells reprogram.