Gene Expression profiles of Bapx1 sorted cells from hindlimbs (HL) of wildtype (WT) and Bapx1 null (HOMO) E12.5 mouse embryos

This study aims to look at gene expresion profiles between wildtype and Bapx1 knockout cells of the hindlimbs in a E12.5 mouse embryo. Instead of looking at the whole hindlimbs,only cells expressing Bapx1 were sorted by Fluroscent Activated Cell Sorting (FACS) and subjected to expression profiling by microarray. EGFP was knocked into the Bapx1 locus by homologous recombination in mESC followed by generation of stable mice line. Wildtype and homozygous embryos expressing EGFP was was used to dissected out the hindlimbs and Fluroscent activated cell sorting was used to sort out cell expressing Bapx1 as marked by EGFP. Total RNA was extracted and hybidized to an Illumina WG6 v2 mouse array.

本研究旨在探究E12.5小鼠胚胎后肢野生型（wildtype）与Bapx1基因敲除（Bapx1 knockout）细胞的基因表达谱差异。相较于直接分析完整后肢，研究人员仅通过荧光激活细胞分选（Fluorescent Activated Cell Sorting, FACS）筛选表达Bapx1的细胞，并采用微阵列技术开展表达谱分析。研究人员先在小鼠胚胎干细胞（mouse embryonic stem cell, mESC）中通过同源重组（homologous recombination）将增强绿色荧光蛋白（enhanced green fluorescent protein, EGFP）敲入Bapx1基因座，随后构建稳定的小鼠品系。取表达EGFP的野生型及纯合子胚胎，解剖分离其后肢，再通过FACS筛选出以EGFP为标记的表达Bapx1的细胞。提取总RNA后，将其与Illumina WG6 v2小鼠基因芯片进行杂交。