Targeting transcription factor binding sites of the interleukin 7 receptor gene using CRISPR/CAS9 genome editing
Çetin, Ronay (2017) Targeting transcription factor binding sites of the interleukin 7 receptor gene using CRISPR/CAS9 genome editing. [Thesis]
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 receptor 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 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 NF-kB binding site in the enhancer and 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 performed chromatin immunoprecipitation (ChIP) experiments to reveal the binding abilities of transcription factors to the IL-7R gene targeted with CRISPR/Cas9. In this study, we identified important transcription factors that control the survival of lymphocytes of the immune system.
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