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Early Myeloid Cell-Specific Expression of the Human Cathepsin G Gene in Transgenic Mice
Jay L. Grisolano, Gary M. Sclar and Timothy J. Ley
Proceedings of the National Academy of Sciences of the United States of America
Vol. 91, No. 19 (Sep. 13, 1994), pp. 8989-8993
Published by: National Academy of Sciences
Stable URL: http://www.jstor.org/stable/2365955
Page Count: 5
You can always find the topics here!Topics: Transgenes, Mice, Bone marrow, Embryos, Myeloid cells, Transgenic animals, Genes, Bone marrow cells, Antibodies, Liver
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The human cathepsin G (CG) gene is expressed only in promyelocytes and encodes a neutral serine protease that is packaged in the azurophil (primary) granules of myeloid cells. To define the cis-acting DNA elements that are responsible for promyelocyte-specific "targeting," we injected a 6-kb transgene containing the entire human CG gene, including coding sequences contained in a 2.7-kb region, ≈2.5 kb of 5' flanking sequence, and ≈0.8 kb of 3' flanking sequence. Seven of seven "transient transgenic" murine embryos revealed human CG expression in the fetal livers at embryonic day 15. Stable transgenic founder lines were created with the same 6-kb fragment; four of five founder lines expressed human CG in the bone marrow. The level of human CG expression was relatively low per gene copy when compared with the endogenous murine CG gene, and expression was integration-site dependent; however, the level of gene expression correlated roughly with gene copy number. The human CG transgene and the endogenous murine CG gene were coordinately expressed in the bone marrow and the spleen. Immunohistochemical analysis of transgenic bone marrow revealed that the human CG protein was expressed exclusively in myeloid cells. Expression of human CG protein was highest in myeloid precursors and declined in mature myeloid cells. These data suggest that the human CG gene was appropriately targeted and developmentally regulated, demonstrating that the cis-acting DNA sequences required for the early myeloid cell-specific expression of human CG are present in this small genomic fragment.
Proceedings of the National Academy of Sciences of the United States of America © 1994 National Academy of Sciences