TARGETING TRANSCRIPTION FACTOR BINDING SITES OF THE INTERLEUKIN 7 RECEPTOR GENE USING CRISPR/CAS9 GENOME EDITING
Molecular Biology, Genetics and Bioengineering, MSc Thesis, 2017
Prof. Batu Erman (Thesis Advisor), Prof. Selim Çetiner,
Asst. Prof. Özgür Gül
Date & Time: 31.07.2017 & 11 AM
Place: FENS G035
Keywords: T lymphocyte, interleukin-7 receptor, CRISPR/Cas9, genome engineering, regulation of transcription
The interleukin 7 cytokine receptor protein (IL-7R) encoded by the Il-7R gene is expressed in immune system cells and it allows these cells to be signaled by the IL-7 survival factor. Mutations in the IL-7R pathway are associated with primary immunodeficiency diseases. Polymorphisms in the IL-7R gene are directly linked to T-cell acute lymphoblastic leukemia (T-ALL) and multiple sclerosis (MS). These findings point to the importance of this signal in health. We have assessed the transcription mechanism of the IL-7R gene in T lymphocytes. We identified the functional significance of transcription factors that bind to the IL-7R gene control elements. To this end, we generated mutations in the IL-7R gene by using CRISPR/Cas9 mediated genome editing. We targeted RORgT and Gfi1 binding sites in intron 2 of the IL-7R gene. We identified the significance of these mutations on IL-7R expression. We set chromatin immunoprecipitation (ChIP) experiment to reveal the binding abilities of transcription factors to the IL-7R gene targeted with CRISPR/Cas9. With these study, we aimed to identify important transcription factors that control the survival of lymphocytes of the immune system.