Deep transcriptome analysis of mouse embryonic fibroblast following simultaneous overexpression of Stella, Oct4 and Nanos2 (SON)

We constructed a polycistronic lentiviral vector to overexpress 3 germ cell specific genes (Stella, Oct4 and Nanos2) in mouse embryonic fibroblast (MEFs) and evaluated the transcriptome portrait in partially reprogrammed cells.We sequenced RNA samples from bulk cell population of two biological duplicates of MEF-GFP (control) and MEF-SON (overexpressed) 21 days post infection. Differential expression analysis of 50 M pair-end read per samples showed overexpression of neurogenesis, blood vessel and proliferation related genes and downregulation of chondroitin sulphate metabolic process, nitric oxide production and innate immune response genes. Overall design: Examination of whole transcriptome following concurrent overexpression of Stella, Oct4 and Nanos2 in MEFs.

本研究构建了多顺反子慢病毒载体，以在小鼠胚胎成纤维细胞（mouse embryonic fibroblast, MEFs）中过表达3个生殖细胞特异性基因（Stella、Oct4及Nanos2），并对部分重编程细胞的转录组全貌进行了评估。我们对感染后21天的MEF-GFP（对照组）与MEF-SON（过表达组）的两份生物学重复的整体细胞群体RNA样本进行了测序，每个样本的双端测序读段量达50M。对每个样本的50M双端测序读段进行差异表达分析后发现，神经发生、血管生成及增殖相关基因的表达量显著上调，而硫酸软骨素代谢过程、一氧化氮生成及先天免疫应答相关基因的表达量则显著下调。实验设计概述：探究在小鼠胚胎成纤维细胞中同时过表达Stella、Oct4及Nanos2后，其全转录组的表达变化。