RNA sequencing of control versus tauR406W transgenic Drosophila heads at day 10 of adulthood
Paired-end RNA sequencing was performed on heads of control and human tauR406W transgenic Drosophila aged to day 10 of adulthood. TauR406W is one of many mutations in the Microtubule-Associated Protein Tau (MAPT) gene that cause an autosomal dominant neurological disorder termed Frontotemporal Lobar Degeneration (FTLD)-tau with MAPT mutation. The tauR406W Drosophila model was developed by Dr. Mel Feany, and has been used widely to study tau biology due to its mild toxicity at day 10 of adulthood, which is convenient for genetic analyses and precedes exponential decline in survival. Using control and tauR406W Drosophila RNA-seq datasets, we found that CREB-regulated genes are overrepresented among differentially expressed genes in tauR406W transgenic Drosophila. Overall design: Control (Elav-Gal4/+) and tauR406W transgenic (Elav-Gal4/+;UAS-tauR406W/+) Drosophila were aged to day 10 of adulthood. RNA was extracted from heads using Trizol. Each biological replicate consisted of 15-30?ng of total RNA from 18 pooled male and female Drosophila heads per genotype. RNA-sequencing was performed on six biologically independent replicates per genotype. The Ovation RNA-Seq System for Drosophila (#0350, NuGen) was used for library preparation according to the User Guide, including depletion of Drosophila rRNAs. After quantification by Qubit and bioanalysis, barcoded libraries were pooled, purified by magnetic bead extraction and sequenced on the Illumina HiSeq 3000 platform with 100 base pair paired-end sequencing.