Expression of the Ly-6E.1 (Sca-1) transgene in adult hematopoietic stem cells and the developing mouse embryo.
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.
Miles
C
C
Laboratory of Gene Structure and Expression, National Institute for Medical Research, London, UK.
Sanchez
M J
MJ
Sinclair
A
A
Dzierzak
E
E
D000818:Animals; D000950:Antigens, Ly / Q000096:biosynthesis* / Q000235:genetics; D016026:Bone Marrow Transplantation; D004622:Embryo, Mammalian; D005314:Embryonic and Fetal Development; D018507:Gene Expression Regulation, Developmental; D017930:Genes, Reporter; D006412:Hematopoietic Stem Cells / Q000378:metabolism*; D008565:Membrane Proteins / Q000096:biosynthesis* / Q000235:genetics; D051379:Mice; D008810:Mice, Inbred C57BL; D008808:Mice, Inbred CBA; D008822:Mice, Transgenic; D011828:Radiation Chimera; D012045:Regulatory Sequences, Nucleic Acid; D014158:Transcription, Genetic; D018183:Transplantation Chimera; D001616:beta-Galactosidase / Q000096:biosynthesis
D000950:Antigens, Ly; C074650:Ly6a protein, mouse; D008565:Membrane Proteins; D001616:beta-Galactosidase