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First published online July 28, 2005
Stem Cells Vol. 23 No. 9 October 2005, pp. 1378 -1388
doi:10.1634/stemcells.2005-0090; www.StemCells.com
© 2005 AlphaMed Press

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Transgenic Analysis of the Stem Cell Leukemia +19 Stem Cell Enhancer in Adult and Embryonic Hematopoietic and Endothelial Cells

Lev Silbersteina, María-José Sánchezb, Merav Socolovskyc, Ying Liuc, Gary Hoffmana, Sarah Kinstona, Sandie Piltza, Mark Bowena, Laure Gambardellad, Anthony R. Greena, Berthold Göttgensa

a Department of Hematology, Cambridge Institute for Medical Research, University of Cambridge, Cambridge, United Kingdom;
b Centro Andaluz de Biologia del Desarrollo, Universidad Pablo de Olavide, Seville, Spain;
c University of Massachusetts Medical School, Department of Pediatrics (Hematology/Oncology Division) and Department of Cancer Biology, Worcester, Massachusetts, USA;
d The Laboratory of Lymphocyte Signaling and Development, The Babraham Institute, Babraham, Cambridge, United Kingdom

Key Words. Hematopoietic stem cell • Transcription factor SCL/tal-1 • Transcriptional regulation • Experimental models

Correspondence: Berthold Göttgens, D.Phil., Department of Hematology, Cambridge Institute for Medical Research, Cambridge University, Hills Road, Cambridge CB2 2XY, U.K. Telephone: 44-1223-336829; Fax: 44-1223-762670; e-mail: bg200{at}cam.ac.uk

Appropriate transcriptional regulation is critical for the biological functions of many key regulatory genes, including the stem cell leukemia (SCL) gene. As part of a systematic dissection of SCL transcriptional regulation, we have previously identified a 5,245-bp SCL +18/19 enhancer that targeted embryonic endothelium together with embryonic and adult hematopoietic progenitors and stem cells (HSCs). This enhancer is proving to be a powerful tool for manipulating hematopoietic progenitors and stem cells, but the design and interpretation of such transgenic studies require a detailed understanding of enhancer activity in vivo. In this study, we demonstrate that the +18/19 enhancer is active in mast cells, megakaryocytes, and adult endothelium. A 644-bp +19 core enhancer exhibited similar temporal and spatial activity to the 5,245-bp +18/19 fragment both during development and in adult mice. Unlike the +18/19 enhancer, the +19 core enhancer was only active in adult mice when linked to the eukaryotic reporter gene human placental alkaline phosphatase. Activity of a single core enhancer in HSCs, endothelium, mast cells, and megakaryocytes suggests possible overlaps in their respective transcriptional programs. Moreover, activity in a proportion of thymocytes and other SCL-negative cell types suggests the existence of a silencer elsewhere in the SCL locus.




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