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The purpose of the project is to knock out the Wfs1 gene in Drosophila (fruit fly) cells and test the hypothesis that this gene is involved in regulating intracellular calcium levels. To test this hypothesis, the gene will first be knocked out using CRISPR-Cas9 molecular tools that are readily available in Dr. Shanle's lab. This will effectively inactivate the Wsf1 gene in cells. To do this, we will transfect Drosophila cells with CRISPR-Cas9 targeting the gene. Next, we will confirm that the gene is mutated using genomic DNA purification, polymerase chain reaction, and sequencing. Finally, we will treat cells with ionomycin to increase intracellular calcium levels and measure cell death. The results for Wfs1 knockout cells will be compared to wild type cells with normal Wfs1. We hypothesize that cells lacking Wis1 will be more sensitive to ionomycin because they cannot properly transport calcium from the endoplasmic reticulum.


Independent Research, Faculty Advisor: Dr. Erin Shale

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