RNAseq Analysis of Transcriptome expression profiles in E13.5 Foxf2-/-Osr2RFP/+ and control palatal mesenchymal cells

Purpose: The purpose of this study is to compare the transcriptome expression profiles of E13.5 Foxf2-/-;Osr2RFP/+ and control palatal mesenchyme by using RNA-seq analysis. Methods: Foxf2+/- female mice were crossed with Foxf2+/-;Osr2RFP/+ male mice.The embryos were harvested at E13.5. The pair of palatal shelves were dissected from each Osr2-RFP+ embryo. The RFP+ palatal mesenchyme cells were isolated by using fluorescence-activated cell sorting (FACS). RNA-seq analysis was carried out using the FACS-isolated palatal mesenchyme from Foxf2-/-;Osr2RFP/+, Foxf2+/-;Osr2RFP/+ and Osr2RFP/+embryos, respectively. Overall design: The transcriptome expression profiles of E13.5 control and Foxf2-/-Osr2RFP/+ palatal mesenchyme by using RNA-seq analysis, in duplicates, using Illumina HisEq 2000.

研究目的：本研究拟通过RNA测序（RNA-seq）分析，对比胚胎发育第13.5天（E13.5）Foxf2基因敲除纯合且Osr2标记RFP阳性（Foxf2-/-;Osr2RFP/+）小鼠与对照组腭间充质的转录组表达谱。
实验方法：将Foxf2杂合敲除雌性小鼠与Foxf2杂合敲除且Osr2-RFP阳性雄性小鼠进行杂交。于胚胎发育第13.5天（E13.5）收取胚胎，从每枚Osr2-RFP阳性胚胎中分离双侧腭突，通过荧光激活细胞分选（fluorescence-activated cell sorting）富集RFP阳性的腭间充质细胞。分别以Foxf2-/-;Osr2RFP/+、Foxf2+/-;Osr2RFP/+及Osr2RFP/+胚胎的FACS分选腭间充质细胞开展RNA-seq分析。
整体实验设计：采用Illumina HisEq 2000测序平台，对E13.5天对照组与Foxf2-/-;Osr2RFP/+小鼠的腭间充质转录组表达谱进行RNA-seq分析，设置两份生物学重复。