Molecular Biology and Genetics MS Thesis Defense by Fatma Sadife İşleyen

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KOÇ UNIVERSITY

GRADUATE SCHOOL OF SCIENCES & ENGINEERING

MOLECULAR BIOLOGY AND GENETICS

MS THESIS DEFENSE BY FATMA SADİFE İŞLEYEN

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Title: Transcriptional Control of Stim1 gene by Transcription Factor NEUROD2

Speaker: Fatma Sadife İşleyen

Time: June 19, 2017, 10:00

Place: SCI 103

Koç University

Rumeli Feneri Yolu

Sariyer, Istanbul

Thesis Committee Members:

Dr. Gülayşe İnce Dunn (Advisor, Koc University)

Prof. Dr. Halil Kavaklı  (Koc University)

Dr. Umut Şahin (Boğaziçi University)

Abstract:

Neurogenic differentiation factor 2 (NEUROD2) transcription factor, a member of basic helix-loop-helix transcription factor family, have essential roles in brain development. Studies performed in our laboratory have revealed Stim1 gene among the top targets of NEUROD2. STIM1 functions as a sensor of endoplasmic reticulum (ER) calcium store levels. Upon store depletion, STIM1 activates store-operated calcium entry (SOCE). NEUROD2 binds to an intronic element within Stim1 gene and regulates its expression. My thesis is composed of three major parts. In the first part, I investigated the functions of NEUROD2 by binding to Stim1 intronic element. I found that NEUROD2 does not function as a repressor by binding to Stim1 intronic element. Indeed, my results suggest that NEUROD2 might function as part of an insulator complex. In the second part, I investigated the effect of NEUROD2 on SOCE in cortical neurons by calcium imaging. My results indicated that the overexpression of Neurod2 does not cause a change in neuronal SOCE response at 5 day in vitro (5DIV). I also tested the effect of Neurod2 knock-down on neuronal SOCE response at 5DIV but my results were inconclusive. In the last part, I aimed to delete Stim1 intronic region comprising clustered E-boxes where NEUROD2 binds to by using CRISPR-Cas9 genome editing system. A better understanding of functions of NEUROD2 regulation of Stim1 expression can be developed by examining the effects of this deletion in the context of cortical development.