Medulloblastoma is the most common malignant pediatric cancer affecting the central nervous system. This malignancy targets the cerebellum and its tendency to metastasize makes non-invasive treatment difficult. Previously, our lab has identified ARHGAP36, a RAS homolog GTPase activating protein, as a potential driver of medulloblastoma in a Sleeping Beauty insertional mutagenesis screen for this disease. Here, we investigated ARHGAP36’s oncogenic potential by designing a knockout vector to alter ARHGAP36 expression in the human medulloblastoma cell line, Ons76. Since ARHGAP36 is a suspected oncogene, we hypothesize deleting ARHGAP36 will lower proliferation and inhibit anchorage independence in Ons76. In this study, we were able to successfully design and construct a CRISPR/Cas9 knockout vector that was designed to create a large deletion within the ARHGAP36 endogenous gene. We achieved alterations in both alleles of the gene and studied its expression. We observed lowered ARHGAP36 protein expression in our treated samples compared to wild type expression using Western blotting. Recognizing ARHGAP36’s oncogenic potential may aid in future research for less invasive therapies for medulloblastoma.
This research was supported by the Undergraduate Research Opportunities Program (UROP).
Larsson, Alex T.; Jackson, Pauline J.; Largaespada, David A..
CRISPR-Mediated Alteration of ARHGAP36 Expression in Human Medulloblastoma.
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