Mentor: Paul James, Ph.D.

Sodium Hydrogen Exchangers (NHEs) are a family of proteins that utilize the electrochemical gradient of Na+ influx to export intracellular H+ ions1. The NHE10 protein, which has its expression localized to the principal piece of sperm, has been shown to be essential for sperm motility2. Mice with the NHE10 encoding gene, SLC9C1, knocked-out are completely infertile due to immotile sperm. The tissue specific expression of NHE10 may be due to a variety of potential epigenetic factors. One of these, DNA methylation, is an epigenetic mechanism involving the addition of a methyl group to a Cytosine in a CpG dinucleotide. This DNA feature has been implicated to have significant regulatory effects on gene expression. Our lab has identified the cell line HeLa as having a highly methylated SLC9C1 promoter region. After expression analysis, we revealed that this cell line has low expression of this gene. We investigated if demethylating the promoter region of the SLC9C1 gene can increase its expression. We plan to use a dCas9 fused to a Ten-eleven translocation methylcytosine dioxygenase 1 (TET1) guided by a designed sgRNA to achieve targeted demethylation.

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