Miles C, Sanchez M J, Sinclair A, Dzierzak E
Laboratory of Gene Structure and Expression, National Institute for Medical Research, London, UK.
Development. 1997 Jan;124(2):537-47. doi: 10.1242/dev.124.2.537.
The mouse hematopoietic marker Sca-1, encoded by the Ly-6E.1 and Ly-6A.2 genes, has been instrumental in the enrichment and characterization of the stem cell for the adult blood system. In the studies reported here, we use Ly-6E.1 genomic fragments to direct expression of a lacZ marker transgene in vivo to study Ly-6E.1 specific regulatory elements in the hematopoietic stem cell and to localize these cells in the developing mouse embryo. We demonstrate that a region approximately 9 kb downstream from the transcriptional start site is required for the distinct, restricted expression pattern of the Ly-6E.1-lacZ transgene within adult hematopoietic stem cells and embryos. We also demonstrate that viable and functional lacZ-expressing hematopoietic stem cells can be enriched by FDG staining and flow cytometric sorting. The Ly-6E.1-lacZ-mediated enrichment of hematopoietic stem cells from adult transgenic bone marrow in combination with the temporal expression pattern of the transgene in the pro/mesonephros suggest an intraembryonic site of development for these cells in the mouse.
由Ly-6E.1和Ly-6A.2基因编码的小鼠造血标志物Sca-1,在成体血液系统干细胞的富集和特性鉴定中发挥了重要作用。在本文报道的研究中,我们使用Ly-6E.1基因组片段在体内指导lacZ标记转基因的表达,以研究造血干细胞中Ly-6E.1特异性调控元件,并在发育中的小鼠胚胎中定位这些细胞。我们证明,转录起始位点下游约9 kb的区域是Ly-6E.1-lacZ转基因在成体造血干细胞和胚胎中独特、受限表达模式所必需的。我们还证明,通过FDG染色和流式细胞术分选可以富集有活力且功能正常的表达lacZ的造血干细胞。Ly-6E.1-lacZ介导的从成年转基因骨髓中富集造血干细胞,结合转基因在前/中肾中的时间表达模式,提示了这些细胞在小鼠胚胎内的发育位点。