Gene expression profiles of E13.5 developing podocyte in the developing kidney isolated from MafB-GFP transgenic mice using FACS on 1.0 ST Array chip. (GUDMAP Series ID: 32). Mus musculus

The long term objective is to create an encyclopedia of the expression levels of all genes in multiple components of the developing kidney. The central thesis is straightforward. The combination of fluorescent activated cell sorting (FACS) plus microarray analysis offers a powerful, efficient and effective method for the creation of a global gene expression atlas of the developing kidney. Microarrays with essentially complete genome coverage can be used to quantitate expression levels of every gene in FACS isolated components of the developing kidney. The ensuing rapid read-out provides an expression atlas that is more sensitive, more economical and more complete than would be possible by in situ hybridizations alone. Overall design: MafB-GFP BAC transgenic mice were utilized to isolate the podocyte cells from the developing embryonic kidneys. Podocyte cells were isolated from the kidney using trypsinization and FACS. RNA was isolated and the gene expression profiles were determined by microarrays.

本数据集的长期研究目标为构建发育中肾脏多组分的全基因表达谱百科全书。其核心论点清晰明确：荧光激活细胞分选（fluorescent activated cell sorting, FACS）联合微阵列分析，可为发育肾脏的全局基因表达图谱构建提供一种高效、稳健且可靠的技术手段。覆盖度近乎完整的微阵列，可用于定量分析经FACS分离得到的发育肾脏组分中所有基因的表达水平。由此获得的快速检测结果，可生成比单纯原位杂交（in situ hybridization）技术更为灵敏、经济且完整的基因表达图谱。

总体实验设计：本研究采用MafB-GFP细菌人工染色体（bacterial artificial chromosome, BAC）转基因小鼠，从胚胎发育阶段的肾脏中分离足细胞。通过胰酶消化结合FACS从肾脏组织中分离足细胞。提取总RNA后，通过微阵列分析测定其基因表达谱。