Transcriptomics profiling of M6A RNA modifications in tnf-alpha induced apoptosis için kapak resmi
Transcriptomics profiling of M6A RNA modifications in tnf-alpha induced apoptosis
Akçaöz, Azime, author.
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Fiziksel Tanımlama:
viii, 58 leaves: charts;+ 1 computer laser optical disc.
Apoptosis is a form of programmed cell death that occurs as a result of physiological or pathological causes. TNF-alpha, which has a regulatory role in immune system cells, stimulates apoptosis through the external pathway. For this reason, it can be used for the treatment of various diseases. Although there are many studies on the regulatory mechanisms of TNF-alpha mediated apoptosis, the contribution of RNA modifications has not been fully elucidated. Regarding the potential role of m6A RNA modification in apoptosis, studies have focused on the effects of regulatory proteins and there is no genomewide m6A methylation profile yet. In the present thesis, firstly, the gene expression patterns of m6A writer, eraser, and reader were examined in HeLa cells and 632 genes with differential m6A methylation pattern were identified by the miCLIP method. 99 genes involved in apoptotic pathways were determined by GO analysis. Candidates were selected based on m6A methylation fold change, intracellular expression level and apoptotic role of the relevant gene. Methylation points in IGV were confirmed and specific validation experiments were performed on these m6A points. SELECT based validation studies showed 1-2 cycle increase in the TNF-alpha group compared to the control group. This confirms the miCLIP data, which also pointed to an increase in m6A methylation. To elucidate the fate of candidate RNAs, the gene expression levels, and translational status of candidate genes were analyzed. METTL3 KD HeLa cells exposed to TNF-alpha exhibited an increase in the expression of PHLDA1, IFI6 and HRK by almost 2-fold. Polysome fractionation assay showed that translation level decreased in TNF-alpha treated METTL3 KD HeLa cells. As a conclusion, global m6A level affected RNA abundance as well as translation.
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Yazar Ek Girişi:
Tek Biçim Eser Adı:
Thesis (Master)--İzmir Institute of Technology:Molecular Biology and Genetics.

İzmir Institute of Technology: Molekular Biology and Genetics --Thesis (Master).
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Tez T002416 QH671 .A31 2021

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