Generation of Knockout Mutants for the tao Gene in D. melanogaster Using the CRISPR/Cas9 Technique and Screening of Gene Function in Border Cell Migration

Deb, Shilpak (2023) Generation of Knockout Mutants for the tao Gene in D. melanogaster Using the CRISPR/Cas9 Technique and Screening of Gene Function in Border Cell Migration. Masters thesis, Indian Institute of Science Education and Research Kolkata.

Text (MS dissertation of Shilpak Deb (18MS021)) Thesis_18MS021.pdf - Submitted Version Restricted to Repository staff only Download (2MB)

Abstract

Gene editing techniques constitute an integral part of genetic research. The CRISPR/Cas9 technique is a relatively recent genome editing method that has been adapted from the bacterial immune system and boasts of greater speed and efficiency compared to its predecessors. In the first part of my project, I employed the CRISPR/Cas9 technique to generate knockout mutants for the tao gene in Drosophila melanogaster. This was done mainly due to the relative unavailability of mutant stocks for tao gene that is known to play a significant role in the epithelial morphogenesis in the Drosophila egg chamber. I have used a single-target guide RNA for generating the mutants in question and based on the lethality observed, I speculate that I have been successful in creating a single mutant line. For the second part of my project, I have conducted a separate unrelated study to observe the effects of a set of genes (tsc1, nedd8, lnc rox2, ben, sarm, vps13, gig, rb, e(bx) and ter94) on the border cell migration phenomenon during oogenesis in D. melanogaster. This was done to validate previous speculations that these genes may play a role in the migration of border cells. However, on downregulating the expression of the mentioned genes in the anterior follicle cells and the border cell cluster, I have not noticed any significant defect in the migration process.

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