Investigating the transcriptional control of the MECP2 gene by using a newly developed cell line
| dc.contributor.author | Lockman, Sandhini | |
| dc.contributor.examiningcommittee | Merz, David (Biochemistry & Medical Genetics) | en_US |
| dc.contributor.examiningcommittee | Leygue, Etienne (Biochemistry & Medical Genetics) | en_US |
| dc.contributor.supervisor | Rastegar, Mojgan | |
| dc.date.accessioned | 2023-04-24T15:56:22Z | |
| dc.date.available | 2023-04-24T15:56:22Z | |
| dc.date.copyright | 2023-04-21 | |
| dc.date.issued | 2023-03-29 | |
| dc.date.submitted | 2023-04-21T19:37:02Z | en_US |
| dc.degree.discipline | Biochemistry and Medical Genetics | en_US |
| dc.degree.level | Master of Science (M.Sc.) | en_US |
| dc.description.abstract | Neuronal DNA methylation is critical to neuronal cell development, maturation, and function. This was illustrated- in large part- by the discovery of Methyl CpG binding protein 2 (MeCP2) as a reader of DNA methylation in the brain. If this role is impaired in neurons, as is seen in individuals diagnosed with Rett Syndrome (RTT), severe mental disability ensues. A contributing factor to this level of severity could be attributed to the high level of MeCP2 observed in mature neurons, fundamental for normal development in terms of facilitating synaptogenesis and dendrite formation. For normal function of neurons it is also important that MeCP2 splice variants (E1 and E2) maintain their expression levels within a certain threshold. MeCP2 protein and transcript levels vary depending on cell type and organ system in which they reside. In the human brain, various anatomical components possess varying levels of MECP2 transcripts. Interestingly, across both MECP2E1 and E2 transcripts, the expression was highest in the human cerebellum out of four brain regions surveyed including the cortex, amygdala, and hippocampus. In this study, a cell model was created using a proliferative cell line originating from the cerebellum. A stably transduced cell line was created providing a reproducible screening assay for which to quantify Mecp2 promoter activity under various conditions. Two different gene reporter assays were used in the same cell lines in order to investigate the function of the MeCP2 isoforms on the promoter elements. Additional reporter assays were conducted to determine the effects of a potential therapeutic drug that targets metabolic pathways that are deregulated in RTT patients. This study provides characterization of the processes taking place at the promoter by quantifying promoter activity levels using a validated and reliable brain cell model. It also serves as a foundation to identify areas for further investigation into the molecular mechanisms of MeCP2. | en_US |
| dc.description.note | May 2023 | en_US |
| dc.description.sponsorship | Ontario Rett Syndrome | en_US |
| dc.identifier.uri | http://hdl.handle.net/1993/37288 | |
| dc.language.iso | eng | en_US |
| dc.rights | open access | en_US |
| dc.subject | MeCP2 | en_US |
| dc.subject | Rett Syndrome | en_US |
| dc.subject | Metformin | en_US |
| dc.subject | Daoy cells | en_US |
| dc.subject | Epigenetics | en_US |
| dc.subject | Transcription | en_US |
| dc.title | Investigating the transcriptional control of the MECP2 gene by using a newly developed cell line | en_US |
| dc.type | master thesis | en_US |
| local.subject.manitoba | no | en_US |
| project.funder.name | Natural Sciences and Engineering Research Council of Canada | en_US |