Developing a High-throughput Method of Generating Pooled CRISPR-induced Deletions in Drosophila
The CRISPR-Cas9 system has opened doors to our understanding of genome editing, genetic disease, and targeted therapies. This technology has been applied in fruit flies, Drosophila, to generate targeted germline deletions to study genes and chromosome organization. These experiments are currently limited by extensively long procedures, where generating a pool of Drosophila with unique CRISPR deletions requires separate cycles of plasmid cloning of guide RNA, embryo injection, and sequence analysis for each locus. This imposes a restricting upper limit on the number of deletions that can be induced and analyzed at once. My project seeks to develop a high-throughput method for generating precise deletions in flies by modifying this procedure and collapsing these cycles into a multiplexed, four-step approach. This technology will ideally enable systemic generation of a pool of Drosophila mutants, pushing this upper limit to potential ten times the original efficiency or more. This would open possibilities to previously unfeasible large-scale experiments investigating gene regulation and create a valuable tool for Drosophila gene editing.
Message to Sponsor
- Major: Molecular and Cell Biology: Developmental Genetics
- Sponsor: Rose Hill Foundation
- Mentor: Michael Eisen