Determining 4D Nuclear Architecture Dynamics of Ligand-Dependent Transcription Activation Using Live Imaging Techniques
This dissertation discusses the 4D chromosomal dynamics relative to subnuclear organelles and transcription hubs during transcription activation, in an estrogen-dependent system. The phenomenon of transcriptional bursting has been observed across numerous model organisms, where gene transcription does not occur continuously over time but rather in periods of high levels followed by periods of inactivity. I aim to explore such phenomenon in our estrogen inducible model and attempt to decipher the spatial and temporal dynamics associated with transcription burst by using RNA and DNA live cell imaging techniques. In the first chapter, I introduce the concept of burst transcription and relevant literature associated with bursting and transcription regulation. In the second chapter, I have included a review I co-authored on enhancers and its biological and biophysical properties that impact transcription regulation. Chapter 3 is where I present the core of my graduate research work involving live cell visualization techniques. Lastly, Chapter 4 is another paper I assisted on, where we examined how acute E2 enhancer activation results in eRNA-mediated phase-separated condensates that allow for cooperative activation of the most robust ERα enhancers.