Generating a Knockout of CG7709 Gene by Employing CRISPR/Cas9 to Study the Effect on Drosophila Oogenesis and Investigating the Role of Gap Junction Proteins in Specialized Follicle Cells of Drosophila Egg Chamber

China, Arkaprabha (2021) Generating a Knockout of CG7709 Gene by Employing CRISPR/Cas9 to Study the Effect on Drosophila Oogenesis and Investigating the Role of Gap Junction Proteins in Specialized Follicle Cells of Drosophila Egg Chamber. Masters thesis, Indian Institute of Science Education and Research Kolkata.

Text (MS Dissertation of Arkaprabha China (16MS103)) 16MS103_Thesis_file.pdf - Submitted Version Restricted to Repository staff only Download (1MB)

Abstract

Genome editing techniques are an integral part of research to study the function of a certain gene. CRISPR/Cas9 is a relatively recent advancement in genome editing technology that has made it faster and more efficient than the other existing methods. CRISPR/Cas9 was first identified as defense machinery in bacteria which allowed them to detect and eliminate viral nucleic acids present during infection. In the first part of my project, I employed CRISPR/Cas9 technique to knock out a gene of interest, CG7709 or muc91C, from the genome of Drosophila. This is primarily being done to confirm previous observations from RNAi data and elucidate the effects of the muc91C Knockout condition. I have used a dual-target approach to generate a deletion of muc91C. I have been successful in creating several mutant alleles that have been sequenced to precisely map the mutation and have been further validated by Reverse Transcription-PCR. I have further formed a genetic characterization of the muc91C mutant to study the corresponding phenotype effect to understand the function of the gene during Drosophila oogenesis. For a separate unrelated study, in the second part of my project, I have employed Gal4/UAS system to downregulate Innexin channel proteins in the polar cells and the follicle cells to study the probable role of innexins during border cell migration in Drosophila oogenesis.

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