Analysis of mir-17~92 targets in motor neurons. Mus musculus

To identify mir-17~92 targets in MNs, a set of target candidate genes from the intersection of upregulated genes found only in the mir-17~92 KO MNs that were predicted to be direct targets of mir-17~92 by TargetScan. Overall design: Motor neurons in the spinal cord from WT ; Hb9::GFP and mir-17~92 -/- ; Hb9::GFP embryos at E12.5 were isolated by FACS. Subsequently, both WT and KO motor neurons (GFPon) and non motor neurons (GFPoff) transcriptome were analyzed by Agilent SurePrint G3 Mouse GE 8x60K Microarray (Agilent-G4858A-028005).

为鉴定运动神经元（Motor Neurons, MNs）内miR-17~92的靶基因，研究人员从仅在miR-17~92敲除（Knockout, KO）运动神经元中上调的基因交集之中，筛选得到经TargetScan预测为miR-17~92直接靶标的候选靶基因集。实验整体设计：收集胚胎发育第12.5天（E12.5）的野生型（Wild Type, WT）Hb9::GFP纯合胚胎与miR-17~92敲除纯合（mir-17~92 -/- ; Hb9::GFP）胚胎的脊髓组织，通过荧光激活细胞分选（Fluorescence-Activated Cell Sorting, FACS）分离其中的运动神经元。后续采用安捷伦SurePrint G3小鼠全基因组8×60K表达芯片（Agilent SurePrint G3 Mouse GE 8x60K Microarray, 货号Agilent-G4858A-028005），对野生型与敲除型运动神经元（GFP阳性，GFPon）以及非运动神经元（GFP阴性，GFPoff）的转录组进行芯片分析。