Simon M C, Pevny L, Wiles M V, Keller G, Costantini F, Orkin S H
Division of Hematology-Oncology, Children's Hospital, Boston, Massachusetts.
Nat Genet. 1992 May;1(2):92-8. doi: 10.1038/ng0592-92.
Development of definitive (fetal liver-derived) red cells is blocked by a targeted mutation in the gene encoding the transcription factor GATA-1. We used in vitro differentiation of GATA-1- mouse embryonic stem (ES) cells to reveal a requirement for GATA-1 during primitive (yolk sac-derived) erythropoiesis and to establish a rescue assay. We show that the block to development includes primitive, as well as definitive, erythroid cells and is complete at the level of globin RNA expression; that the introduction of a normal GATA-1 gene restores developmental potential both in vivo and in vitro; and that efficient rescue is dependent on a putative autoregulatory GATA-motif in the distal promoter. Use of in vitro differentiated ES cells bridges a gap between conventional approaches to gene function in cell lines and analysis of loss of function mutations in the whole animal.
编码转录因子GATA-1的基因发生靶向突变会阻碍成熟（胎儿肝脏来源）红细胞的发育。我们利用GATA-1基因敲除小鼠胚胎干细胞的体外分化来揭示原始（卵黄囊来源）红细胞生成过程中对GATA-1的需求，并建立了一种拯救试验。我们发现，发育阻断包括原始红细胞和成熟红细胞，并且在珠蛋白RNA表达水平上是完全的；正常GATA-1基因的导入可在体内和体外恢复发育潜能；高效拯救依赖于远端启动子中一个假定的自调控GATA基序。使用体外分化的胚胎干细胞弥合了细胞系中基因功能的传统研究方法与全动物功能缺失突变分析之间的差距。
