Generation of a deletion of the draper gene in Drosophila melanogaster using CRISPR/Cas9
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Abstract
Cells within the body undergo death on a daily basis and those dead cells are constantly being removed so as to prevent accumulation of dead cells that can further cause negative effects. In Drosophila, an important gene necessary for clearance of dead cells is draper. This gene encodes a phagocytic receptor essential for clearance of dead cells. This project explored using CRISPR/Cas9 to edit the Drosophila genome to delete draper to create a mutant. Guide RNAs were designed to produce DNA breakpoints 4.5 kB apart, flanking most of the draper coding region. Oligonucleotides with the guide RNA sequences were cloned into an expression vector, and the vectors were injected into Cas9-expressing flies. Candidate mutants were dissected and their ovaries were analyzed for defective cell clearance. Two lines were shown to have defective cell clearance in the ovary and are potential new deletion alleles of draper.