Single cell RNA sequencing of peripheral retina in control and FGF signaling mutant mouse embryonic tissue

The goal of the study is to identify changes in gene expression and cell distribution in the ciliary margin upon loss of FGF signaing. Neural retinal cells from embryonic day E14 were isolated from control and mutant eyes. Dissociated cells were sorted based on GFP and TdTomato reporter expression in the control and mutant retina respectively. Flow-sorted single cells were processed by the core facility at Columbia University on a 10x platform. 3 control and 3 mutant embryos were pooled and sorted based on reporter expression for single cell sequencing本研究旨在探究成纤维细胞生长因子（FGF）信号通路缺失后，睫状缘（ciliary margin）内基因表达与细胞分布的改变。研究人员从对照组与突变体胚胎的眼球中分离得到胚胎第14天（embryonic day E14）的神经视网膜细胞；分别以对照组视网膜内的绿色荧光蛋白（GFP）报告基因、突变体视网膜内的TdTomato荧光蛋白（TdTomato）报告基因的表达情况为依据，对解离后的细胞进行分选。经流式分选得到的单个细胞由哥伦比亚大学核心实验平台依托10x平台（10x platform）完成处理；将3个对照组胚胎与3个突变体胚胎的样本混合后，基于报告基因表达完成分选，用于单细胞测序。