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An evolutionary study of gurken and its effects on axis formation in Drosophila
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Sottolano, Christopher James. An evolutionary study of gurken and its effects on axis formation in Drosophila. Retrieved from https://doi.org/doi:10.7282/t3-fftx-nn74

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TitleAn evolutionary study of gurken and its effects on axis formation in Drosophila
NameSottolano, Christopher James (author); Yakoby, Nir (chair); Piccoli, Benedetto (member); Geneva, Anthony (member); Kulathinal, Robert (member); Rutgers University; Graduate School - Camden
Date Created2023
Other Date2023-05 (degree)
SubjectDevelopmental biology, Biology, Morphology, Cell signalling, CRISPR, Drosophila, EGFR, Evolution, Gurken
Extent142 pages : illustrations
DescriptionAnimal development depends on combinations of interacting signaling pathways which spatiotemporally regulate gene expression in specific domains of cells, leading to tissue differentiation and organ formation. One such pathway is the epidermal growth factor receptor pathway (EGFR), which is activated during Drosophila oogenesis by TGFα-like ligand Gurken (GRK). This activation leads to establishment of the anterior-posterior and dorsal-ventral axes of the developing fly. EGFR activation controls gene expression in a monolayer of follicle cells surrounding the growing oocyte, which will subsequently give rise to eggshell morphologies. Although GRK is necessary for axial development, orthologs show surprisingly low conservation within the Drosophila genus. Here, we investigate how a system allows for evolutionary changes to an essential ligand while still efficiently functioning to establish oocyte polarity. First, evolutionary changes to cis-regulation of grk mRNA was examined via single component transgenic swap and found that evolution to the 3’UTR hampers efficient translation. Next, we studied GRK protein evolution through the CRISPR/Cas9-mediated deletion of a previously uncharacterized histidine-rich domain. While this domain is not necessary, it contributes substantially to axis formation. Lastly, we generated tools for transgenic experiments in D. nebulosa by assembling a high-quality annotated genome from PacBio long-read sequencing. Employing CRISPR/Cas9, we to successfully disrupted the white gene as proof-of-concept. Overall, this research takes advantage of computational methods, including sequence and image analysis, to predict and model evolutionary changes. Computational analysis was subsequently subjected to experimental validation through genome engineering and molecular assays, to study how the ligand’s path to EGFR signaling evolved across species.
NotePh.D.
NoteIncludes bibliographical references
NoteIncludes vita
Genretheses
Persistent URLhttps://doi.org/doi:10.7282/t3-fftx-nn74
LanguageEnglish
CollectionCamden Graduate School Electronic Theses and Dissertations
Organization NameRutgers, The State University of New Jersey
RightsThe author owns the copyright to this work.
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