Michael Hilzendeger

Program: Cancer Biology

Current advisor: Charles Kaufman, MD, PhD

Undergraduate university: University of Virginia, 2019

Enrollment year: 2021

Research summary
Engineering refined models of tumorigenesis in zebrafish using prime editing

Zebrafish serve as a versatile model of human disease. Historically, due to the difficult nature of genome editing in zebrafish, models mainly use transgenesis to express mutant versions of genes of interest. While transgenesis is advantageous in many cases, experimental variables such as transgene expression can vary significantly between experiments, reducing the tractability and reliability of these models. Within the last decade, CRISPR and now prime editing have revolutionized the field of disease modeling by allowing for the introduction of mutations into the genome with great specificity and efficiency. Here, we use a modified prime editing system with the goal of introducing oncogenic mutations at endogenous loci in the zebrafish genome, focusing on the braf locus. Melanoma is modeled in zebrafish using a transgenic construct that overexpresses human BRAF V600E, one of the most common driver mutations found in cutaneous melanoma, under a melanocyte-specific promoter. Through modeling melanoma using mutations at the endogenous braf locus, we hope to further refine our understanding of melanoma initiation and progression by decoupling ectopic oncogenic braf expression from the exogenous mitf promoter. In the future, our zebrafish model of melanoma can be further refined by creating a tissue-specific prime editing system, as has been done for Cas9 based mutagenesis, as well as investigate other oncogenic mutations frequently found in melanoma and other cancers.

Graduate publications