Multifunctional reversible knockout/reporter system enabling fully functional reconstitution of the AML1/Runx1 locus and rescue of hematopoiesis
Multifunctional reversible knockout/reporter system enabling fully functional reconstitution of the AML1/Runx1 locus and rescue of hematopoiesis
复制标题
DOI：
10.1002/gene.20190
复制
发表时间：
2006-03-01
期刊：
影响因子：
1.5
通讯作者：
Medvinsky, A
中科院分区：
文献类型：
--
作者：
Samokhvalov, IM;Thomson, AM;Medvinsky, A
缺陷于runt同源结构域转录因子Runx 1的小鼠在胚胎第12.5天死于子宫内严重贫血。通过靶向插入loxP-侧翼的多部分基因终止/陷阱盒产生可再活化的Runx 1敲除胚胎干细胞（ESC）和小鼠系统，所述多部分基因终止/陷阱盒设计成同时消除Runx 1的表达并报告其启动子的活性。该盒的框内LacZ报告基因使近端和远端启动子的活性能够被差异监测。尽管Runx 1缺失的胚胎干细胞能够在体外分化为原始红细胞，但它们产生粒细胞/巨噬细胞或混合骨髓-红细胞胚状体的能力丧失。Cre介导的再激活恢复了Runx 1结构的完整性，并挽救了ESCs的造血分化潜力。具有重新激活的等位基因的小鼠存活，没有表现出造血缺陷，并且适当地表达Runx 1的所有主要剪接异构体。这种多用途小鼠模型将有助于分析造血发育中的关键Runx 1依赖性检查点。
Mice deficient in the runt homology domain transcription factor Runx1 die of severe anemia in utero by embryonic day (E)12.5. A reactivatable Runx1 knockout embryonic stem cell (ESC) and mouse systems were generated by the targeted insertion of a loxP-flanked multipartite gene stop/trap cassette designed to simultaneously ablate the expression of Runxl and report on the activity of its promoters. The cassette's in-frame LacZ reporter enabled activities of the proximal and the distal promoters to be differentially monitored. Although Runx1-null ESCs were capable of primitive erythroid differentiation in vitro, their capacity to generate granulocyte/macrophage or mixed myelo-erythroid embryoid bodies was lost. Cre-mediated reactivation restored Runx1 structural integrity and rescued the hematopoietic differentiation potential of ESCs. Mice with the reactivated allele survived, showed no hematopoietic deficit, and expressed all major splice isoforms of Runx1 appropriately. This multipurpose mouse model will be useful for the analysis of the critical Runx1-dependent checkpoint(s) in hematopoietic development.