Gata3 directly regulates cell cycle exit-coupled lens fiber cell differentiation in mouse embryonic lens

Gata3 is a DNA-binding transcription factor involved in cellular differentiation in a variety of tissues. To investigate the role of Gata3 in lens development we conducted lens-specific conditional loss-of-function model of Gata3 and confirmed abnormal regulation of cell cycle exit-coupled lens differentiation and retention of nuclei in the mutated lens fibers. We analyzed transcriptomes between control and mutated lenses by RNA-seq and revealed dysregulation of genes encoding critical components of the lens differentiation cascade, including a subset of crystallin genes, down-regulation of Dnase2b and reduced expression of Cdkn1b/p27 and Cdkn1c/p57. Lens mRNA profiles of E14.5 wild-type (WT) and Gata3 lens-specific knockout (CKO) mice were analyzed by RNA-seq. Each sample represents the pool of 10 lenses from 5 embryos and total number of 3 biological replicates was used for transcriptome sequencing using Illumina HiSeq400 system.Gata3是一种DNA结合转录因子，参与多种组织的细胞分化过程。为探究Gata3在晶状体发育中的作用，我们构建了Gata3晶状体特异性条件性功能丧失（conditional loss-of-function）模型，并证实突变的晶状体纤维中存在细胞周期退出耦联的晶状体分化调控异常，以及细胞核滞留现象。我们通过RNA测序（RNA-seq）分析对照组与突变组晶状体的转录组，揭示了晶状体分化级联反应关键编码基因的表达失调，包括一组晶状体蛋白（crystallin）基因、脱氧核糖核酸酶2b（Dnase2b）的下调，以及细胞周期蛋白依赖性激酶抑制剂1b/p27（Cdkn1b/p27）与细胞周期蛋白依赖性激酶抑制剂1c/p57（Cdkn1c/p57）的表达水平降低。本研究采用RNA测序技术，分析了E14.5天野生型（WT）与Gata3晶状体特异性敲除（CKO）小鼠的晶状体mRNA表达谱：每个样本混合5只胚胎的10个晶状体，共设置3次生物学重复，所有样本均通过Illumina HiSeq400系统完成转录组测序。