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Knockout Of Endospanin 1 Via Crispr In Zebrafish, Danio Rerio, Jared Kittinger
Knockout Of Endospanin 1 Via Crispr In Zebrafish, Danio Rerio, Jared Kittinger
Williams Honors College, Honors Research Projects
I made endospanin 1 knockout (KO) zebrafish to examine its effects on lipid and bone metabolism. Endospanin 1, or leptin receptor overlapping transcript (leprot), is a cytosolic protein linked to the protein hormone leptin that influences the trafficking of leptin receptors to the plasma membrane of cells. Genes for endospanin and tyrosinase (a pigmentation enzyme) were targeted via a microinjection of guide RNAs and CRISPR Cas9 into zebrafish embryos at 2-4 cell stages. I was able to disrupt the endospanin 1 gene (based upon the disruption of tyrosinase), but very few mutant zebrafish fully developed into adults. Only low KO …