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Blood, 1 September 2007, Vol. 110, No. 5, pp. 1448-1457. Prepublished online as a Blood First Edition Paper on April 24, 2007; DOI 10.1182/blood-2006-12-060814. This Article Full Text Full Text (PDF) Supplemental Figures All Versions of this Article: blood-2006-12-060814v1 110/5/1448    most recent Alert me when this article is cited Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Rights and Permissions Citing Articles Citing Articles via CrossRef Citing Articles via Google Scholar Google Scholar Articles by Zhang, F. Articles by Thrasher, A. J. Search for Related Content PubMed PubMed Citation Articles by Zhang, F. Articles by Thrasher, A. J. Related Collections Gene Expression Gene Therapy Social Bookmarking                   What's this?  Previous Article  |  Table of Contents  |  Next Article  GENE THERAPY Lentiviral vectors containing an enhancer-less ubiquitously acting chromatin opening element (UCOE) provide highly reproducible and stable transgene expression in hematopoietic cells Fang Zhang1, Susannah I. Thornhill1, Steven J. Howe1, Meera Ulaganathan1, Axel Schambach2, Joanna Sinclair1, Christine Kinnon1, H. Bobby Gaspar1,3, Michael Antoniou4, and Adrian J. Thrasher1,3 1 Centre for Immunodeficiency, Molecular Immunology Unit, Institute of Child Health, University College London, London, United Kingdom; 2 Department of Experimental Hematology, Hannover Medical School, Hannover, Germany; 3 Department of Clinical Immunology, Great Ormond Street Hospital National Health Service (NHS) Trust, London, United Kingdom; 4 Nuclear Biology Group, Department of Medical and Molecular Genetics, Kings College London School of Medicine-Guy's Campus, Guy's Hospital, London, United Kingdom Ubiquitously acting chromatin opening elements (UCOEs) consist of methylation-free CpG islands encompassing dual divergently transcribed promoters of housekeeping genes that have been shown to confer resistance to transcriptional silencing and to produce consistent and stable transgene expression in tissue culture systems. To develop improved strategies for hematopoietic cell gene therapy, we have assessed the potential of the novel human HNRPA2B1-CBX3 UCOE (A2UCOE) within the context of a self-inactivating (SIN) lentiviral vector. Unlike viral promoters, the enhancer-less A2UCOE gave rise to populations of cells that expressed a reporter transgene at a highly reproducible level. The efficiency of expression per vector genome was also markedly increased in vivo compared with vectors incorporating either spleen focus-forming virus (SFFV) or cytomegalovirus (CMV) promoters, suggesting a relative resistance to silencing. Furthermore, an A2UCOE-IL2RG vector fully restored the IL-2 signaling pathway within IL2RG-deficient human cells in vitro and successfully rescued the X-linked severe combined immunodeficiency (SCID-X1) phenotype in a mouse model of this disease. These data indicate that the A2UCOE displays highly reliable transcriptional activity within a lentiviral vector, largely overcoming insertion-site position effects and giving rise to therapeutically relevant levels of gene expression. These properties are achieved in the absence of classic enhancer activity and therefore may confer a high safety profile. CiteULike    Connotea    Del.icio.us    Digg    Reddit    Technorati    What's this?

	Copyright © 2007 by American Society of Hematology         Online ISSN: 1528-0020