Transcriptome profile of Drosophila adult LOF models for motor neuron degenerative disorders. Adult Fly MN-RBP LOF transcriptome

This study addresses the impact of reduced expression of the drosophila orthologues of the human Spinal Muscular Atrophy (SMN1) and Amyotrophic Lateral Sclerosis (FUS and TARDBP) disease genes on the fly neuronal transcriptome. Adult-onset expression of short hairpin (sh) RNA was used to induce RNAi-mediated gene silencing of caz, Smn, TBPH or always early as a control. RNA was prepared from pooled fly heads five to seven days post induction of shRNA expression, quality checked and subjected to RNA-seq following validation of target gene knock-down levels by qPCR. Flies without adult-onset induction of respective shRNA expression served as control. The transcriptome from 3 independent replicates was profiled using Illumina paired-end mRNA-seq.

本研究旨在探究人类脊髓性肌萎缩症（Spinal Muscular Atrophy, SMA，致病基因为SMN1）与肌萎缩侧索硬化症（Amyotrophic Lateral Sclerosis, ALS，致病基因为FUS与TARDBP）的果蝇同源基因表达下调，对果蝇神经元转录组的影响。实验采用成年启动的短发夹RNA（short hairpin RNA, shRNA）诱导RNA干扰（RNAi）介导的基因沉默，分别靶向沉默caz、Smn、TBPH基因，并以always early作为阴性对照。在shRNA诱导表达后的5至7天，收集混合的果蝇头部以提取总RNA，经质量质控后进行RNA测序；同时通过定量聚合酶链式反应（quantitative PCR, qPCR）验证靶基因的敲低效率。未进行成年启动对应shRNA诱导的果蝇作为空白对照。本研究通过3次独立生物学重复，采用Illumina双端mRNA测序技术完成转录组谱分析。