Scimè, AnthonyFine, Kyra Rachel2024-07-182024-07-182024-04-232024-07-18https://hdl.handle.net/10315/42178Myogenic stem cells have the potential to either self-renew or differentiate into muscle fibers. These stem cell fate choices are influenced by metabolic changes which affect the epigenetic landscape, including DNA methylation which alters gene expression patterns. Because we have found that a mitochondrial function of p107, a transcriptional co-repressor, is involved in regulating SC metabolism and fate decisions, we investigated whether p107 influences the mitochondrial DNA methylation signature. For this we used a portable long read sequencer MinION (Nanopore), to sequence and compare the mitochondria DNA methylation pattern of wildtype and p107 genetically deleted (p107KO) C2C12 myogenic cell line. We found that p107KO mitochondrial DNA had decreased methylation levels for OXPHOS genes compared to wild type. Thus, suggesting that p107 might suppress ND4 gene expression. We also compared a novel method of mtDNA isolation to three existing methods in order to determine the most efficient approach for sequencing.Author owns copyright, except where explicitly noted. Please contact the author directly with licensing requests.BiologyBioinformaticsGeneticsElectronic Thesis or Dissertation2024-07-18SequencingMethylationBioinformaticsGeneticsp107C2C12MinIONNanoporeMyoblastsStem cellsMitochondriaEpigeneticsMitogenomeMitoepigenomeMouseMouse geneticsCell cultureDNA isolationLong read sequencingGenomicsOXPHOSOxidative phosphorylationMethylomeDifferential methylation analysisTranscription factors