Scholarly Commons

Determining the relative effectiveness of different RNAi lines at knocking down the hedgehog signaling pathway in various Drosophila tissues

The hedgehog (Hh) signaling pathway has been studied extensively in Drosophila, and is crucial for axonogenesis and patterning events in early embryo development. The Hh protein relieves suppression of transmembrane Smoothed (Smo), resulting in an intracellular cascade that activates Cubitus interruptus (Ci), a transcription factor that regulates Hh target gene expression. Smo RNAi lines were used to inhibit Hh target gene expression. Prior studies using RNAi lines to knock down gene expression in Drosophila have found varying levels of severity in phenotypes across tissues, with short hairpin typically producing the strongest loss-of-function phenotype. Four RNAi lines were used: 2 Valium 20 (short hairpin), 1 VDRC (short hairpin), and 1 Valium 10 (long hairpin). With the variability in effectiveness of RNAi lines across tissues, is unclear which RNAi line will be the most effective at reducing Hedgehog target gene expression in midline glia, but it will likely be a short hairpin line.