RNA-Seq of head tissue from Drosophila melanogaster Wild Type and Adar5G1dAdar-/- mutant. Drosophila melanogaster

Purpose: Validation of Drosophila A-to-I editing sites Methods: We collected heads of 5 day old male dAdar-/- mutant (y, Adar5G1, w)26 and wild type (w1118) flies. Poly(A)+ RNA was used to prepare RNA-seq libraries which were subsequently sequenced single-end by an Illumina GAII Results:We builded a framework to identify RNA editing events using RNA-seq data alone in Drosophila. To validate whether the identified A-to-G sites were bona fide A-to-I editing events, we performed RNA-seq for the D.melanogaster wild-type strain (w1118) and for the Adar5G1 null mutant that eliminates RNA editing. We found that our method achieved high accuracy; 98.2% of all A-to-G sites showed only adenosine in the Adar5G1 sample Conclusions: We anticipate that our method will be very effective in the future to identify RNA editing events in different species. Overall design: mRNA profiles of heads of 5 day old male dAdar-/- mutant (y, Adar5G1, w)26 and wild type (w1118) flies

研究目的：验证果蝇A-to-I编辑（A-to-I editing）位点。
实验方法：我们收集了5日龄雄性dAdar-/-突变体（y, Adar5G1, w）26与野生型（w1118）果蝇的头部组织。采用Poly(A)+ RNA构建RNA测序（RNA-seq）文库，随后通过Illumina GAII测序平台开展单端测序。
研究结果：我们构建了一套仅依托RNA-seq数据即可鉴定果蝇RNA编辑事件的分析框架。为验证所鉴定的A-to-G位点是否为真实的A-to-I编辑事件，我们对黑腹果蝇（D. melanogaster）野生型菌株w1118以及可完全消除RNA编辑的Adar5G1纯合缺失突变体进行了RNA-seq测序。结果显示，本方法具备较高准确性：在Adar5G1突变体样本中，98.2%的A-to-G位点仅检测到腺苷。
研究结论：我们预计本方法在未来可有效应用于不同物种的RNA编辑事件鉴定工作。
整体实验设计：5日龄雄性dAdar-/-突变体（y, Adar5G1, w）26与野生型（w1118）果蝇头部组织的mRNA表达谱