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Question: You hypothesise that EV cargo from aged cells can play a role in changing the epigenetic landscape of stem cells. Outline an experimental design that allows you to explore the influence of EVs on Histone modification. Include a hypothesis, rationale, methodology, what you will quantify and why, and how will you interpret the data in 700 words
. Hypothesis: EVs derived from aged cells can influence the epigenetic landscape of stem cells. Rationale: Epigenetics is important for regulating gene expression and cellular function. As cells age, changes in the epigenome can lead to altered gene expression and decreased function. EVs contain proteins and nucleic acids, including microRNAs, which can alter the epigenetic landscape and impact stem cell development and function. This experiment seeks to examine how EVs from aged cells can influence the epigenetic landscape of stem cells. Methodology: This experiment will use a two-step process to examine the impact of EVs on the epigenetic landscape of stem cells. The first step will be to generate EVs from aged cells. This will involve culturing aged cells in a laboratory setting and collecting EVs derived from the cells using a gradient centrifugation technique. EVs will then be concentrated and tested to assess the relative abundance of specific proteins and microRNAs. The second step will be to examine the effect of EVs on the epigenetic landscape of stem cells. To do this, stem cells will be cultured with or without EVs and then examined for histone methylation, as this is a common epigenetic modification that can influence gene expression. Histone methylation levels will be quantified using a chromatin immunoprecipitation assay. What will be quantified: In this experiment, the relative abundance of proteins and microRNAs in EVs will be quantified, as well as the levels of histone methylation in stem cells. Why: Quantifying EVs will provide a measure of how well the two-step process was able to generate EVs, as well as a measure of the relative abundance of proteins and microRNAs associated with cellular ageing. Quantifying histone methylation levels in stem cells will provide a measure of how exposure to EVs can alter the epigenetic landscape. Interpreting the Data: Data will be analyzed to assess the impact of EVs on histone methylation in stem cells. If exposure to EVs leads to changes in histone methylation, this will indicate that EVs derived from aged cells can influence the epigenetic landscape of stem cells.
Jan. 16, 2023, 4:52 a.m.
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