Date of Award

2011

Degree Type

Thesis

Degree Name

Master of Science

Program

Biochemistry

Supervisor

Dr. Joseph Torchia

Abstract

p/CIP is a transcriptional coactivator that binds liganded nuclear hormone receptors, as well as other transcription factors, and facilitates transcription through direct recruitment of accessory factors. p/CIP is highly expressed in undifferentiated mouse embryonic stem (mES) cells and is downregulated during differentiation. Using siRNA- mediated knockdown of p/CIP in mES cells in combination with microarray analysis I have identified a contingent of essential pluripotency genes which are significantly downregulated including Klf4, Tbx3 and Dax-1. Subsequent chromatin immuno­ précipitation (ChIP) analysis demonstrated that Tbx3 and Dax-1 are direct transcriptional targets of p/CIP. Using the piggyBac transposase system, a mouse ES cell line that expresses Flag-p/CIP in a doxycycline-dependent manner was generated. p/CIP

overexpression increased pluripotency gene expression and promoted the formation of undifferentiated ES cell colonies. Collectively, these results indicate that p/CIP contributes to the maintenance of ES cell pluripotency through direct, as well as indirect, regulation o f essential pluripotency genes.

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