Jessica Caushi '21

Title of Research: Elucidating the Epigenetic Map of the KCC2 Gene.

Mentor: Dr. Alma Rodenas- Ruano 

Abstract: One of the key factors responsible for the homeostatic balance between excitatory and inhibitory synaptic activity is the sudden triggering in the expression of the kcc2 gene, which codes for KCC2, a protein that play a significant role in regulating intracellular chloride levels. My goal for this project was to use zebrafish as an in vivo model to understand how transcriptional mechanisms and epigenetic marks may modify the accessibility of the gene. My main hypothesis is that the activation of kcc2 expression is mediated by epigenetic marks that undergo a shift from a repressive state, where DNA is inaccessible for transcription factor binding, to an activated state, where DNA is made accessible. To test this hypothesis, I first conducted immunostains with synaptotagmin and kcc2 on a zebrafish 5 days post-fertilization, a developmental timeframe where inhibition is already turned on. I then executed a ChIP assay to try and understand what the histone epigenetic map of kcc2 looks like before and after KCC2 expression is triggered. Last, the extraction of RNA and subsequent cDNA synthesis were completed to tell us which genes are expressed and what the expression profile of the genes look like.