Reporters — Deepika Vasudevan

DEVELOPING REPORTERS FOR STUDYING CELLULAR STRESS

Stress response signaling and it’s downstream effects on mRNA translation.

Expression of the 4E-BP-intron-DsRed reporter in adult flies after overnight starvation (fed water only).

The 4E-BP-intron-DsRed reporter shows that many tissues in the fly experiences during development.

Expression of 4E-BP-intron-DsRed, 4E-BP-intron_mutant-DsRed and 4E-BP-upstream-DsRed at the wandering third instar larval stage.

Vector map of the pBC plasmid.

Schematic of the in vivo mono cistronic Drosomycin-GFP reporters (DromP-Drom-GFP, DromP-Tub-GFP).

Over the course of my postdoctoral research, I have generated/characterized several transgenic reporter fly lines that are currently the most facile and sensitive tool for analysis of the ER stress response in Drosophila. These reporters form the the basis for several projects in our laboratory. Here is a partial list of these reporters in rough order of popularity (based on the frequency of reagent requests). The list is best read in conjunction with the signaling pathway schematic in the accompanying figure.

4E-BP-intron-DsRed: 4E-BP intron with multiple ATF4-binding sites driving DsRed
4E-BP-intron_mutant-DsRed: The above reporter with the ATF-binding sites mutated
4E-BP-upstream-DsRed: 4E-BP upstream promoter with FOXO-binding sites driving DsRed
pBiCistronic (pBC): pUAST or pattB with Tubulin promotor backbone with GFP-MCS-RFP
DromP-Drom-GFP: Drosomycin promoter driving Drosomycin 5’UTR-GFP
DromP-Tub-GFP: Drosomycin promoter driving Tubulin 5’UTR-GFP
(PMC5223598, PMC5728446)