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Prepublished online as a Blood First Edition Paper on February 13, 2003; DOI 10.1182/blood-2002-09-2991. This Article Full Text Full Text (PDF) All Versions of this Article: 2002-09-2991v1 101/12/4717    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 HighWire Citing Articles via CrossRef Citing Articles via Google Scholar Google Scholar Articles by Ramezani, A. Articles by Hawley, R. G. Search for Related Content PubMed PubMed Citation Articles by Ramezani, A. Articles by Hawley, R. G. Related Collections Gene Expression Gene Therapy Hematopoiesis Phagocytes Social Bookmarking                   What's this?  Previous Article  |  Table of Contents  |  Next Article  Blood, 15 June 2003, Vol. 101, No. 12, pp. 4717-4724 GENE THERAPY Performance- and safety-enhanced lentiviral vectors containing the human interferon- scaffold attachment region and the chicken -globin insulator Ali Ramezani, Teresa S. Hawley, and Robert G. Hawley From the Hematopoiesis Department, Flow Cytometry Facility, and Blood and Cell Therapy Development, Holland Laboratory, American Red Cross, Rockville, MD; and the Department ofAnatomy and Cell Biology, and Programs in Genetics and Molecular and Cellular Oncology, The George Washington University, Washington, DC. Retroviral vectors are the most efficient means of stable gene delivery to hematopoietic stem cells (HSCs). However, transgene expression from retroviral vectors is frequently subject to the negative influence of chromosomal sequences flanking the site of integration. Toward the development of autonomous transgene expression cassettes, we inserted the human interferon- scaffold attachment region (IFN-SAR) and the chicken -globin 5' DNase I hypersensitive site 4 (5'HS4) insulator both separately and together into a series of self-inactivating (SIN) lentiviral vector backbones. Transduced cells of the human CD34+ hematopoietic progenitor line KG1a—pooled populations as well as individual clones harboring single integrants—were analyzed for reporter expression during culture periods of up to 4 months. Vectors carrying both the 5'HS4 insulator and the IFN-SAR consistently outperformed control vectors without inserts as well as vectors carrying either element alone. The performance of a set of vectors containing the murine stem cell virus long terminal repeat as an internal promoter was subsequently assessed during in vitro monocytic differentiation of transduced primary human CD34+ cord blood cells. Similar to what was observed in the KG1a hematopoietic progenitor cell model, optimal reporter expression in primary monocytes was obtained with the vector bearing both regulatory elements. These findings indicate that the 5'HS4/IFN-SAR combination is particularly effective at maintaining open chromatin domains permissive for high-level transgene expression at early and late stages of hematopoietic development, and thus could be of utility in HSC-directed retroviral vector–mediated gene transfer applications. CiteULike    Connotea    Del.icio.us    Digg    Reddit    Technorati    What's this? This article has been cited by other articles: B. Y. Ryu, M. V. Evans-Galea, J. T. Gray, D. M. Bodine, D. A. Persons, and A. W. 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