i-TRACE RNA interference-based reporter system to distinguish spatiotemporal gene expression in real time versus lineage cells in Drosophila eye
Anuradha Chimata Venkatakrishnan, Paige Elizabeth McBride
Dynamic gene expression pattern along the spatiotemporal axis is a key fundamental feature of all cells. Such changes contribute to cellular responses to stimuli and also execution of highly regulated and sequential developmental programs. During Drosophila (a.k.a fruit fly) eye development, several genes are expressed in a dynamic manner to regulate target genes. Our interest was to understand the exact expression pattern of some of the genes involved in this process. We have used the i-TRACE (RNAi Technique for Real-time And Clonal Expression) system to assess such minute but dynamic changes in expression pattern by combining Gal4/UAS, FLP/FRT, RNAi and fluorescent reporters. Upon crossing with a Gal4 stock, all real time gene expression (Gal4 mediated) is marked by the presence of red fluorescent protein (RFP) and all lineage cells express green fluorescent protein (GFP) independent of Gal4 activity. Additionally, the effect of Gal4 mediated GFP-RNAi expression persists in cells that currently and recently expressed the gene. This enables us to observe even brief changes in expression marked by loss of GFP. Here, we present the expression data of some of the genes that play an important role during Drosophila eye development.
Primary Advisor's Department
Stander Symposium project, College of Arts and Sciences
United Nations Sustainable Development Goals
Good Health and Well-Being
"i-TRACE RNA interference-based reporter system to distinguish spatiotemporal gene expression in real time versus lineage cells in Drosophila eye" (2022). Stander Symposium Projects. 2423.