SEARCH:   Advanced

Prepublished online as a Blood First Edition Paper on February 13, 2003; DOI 10.1182/blood-2002-09-2991. This Article 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 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 Social Bookmarking                   What's this? Submitted October 1, 2002 Accepted February 4, 2003 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* Hematopoiesis Department, American Red Cross Holland Laboratory, Rockville, MD, USA Hematopoiesis Department, American Red Cross Holland Laboratory, Rockville, MD, USA; Flow Cytometry Facility, American Red Cross Holland Laboratory, Rockville, MD, USA Hematopoiesis Department, American Red Cross Holland Laboratory, Rockville, MD, USA; Blood and Cell Therapy Development, American Red Cross Holland Laboratory, Rockville, MD, USA; Department of Anatomy and Cell Biology, and Programs in Genetics and Molecular and Cellular Oncology, The George Washington University, Washington, DC, USA * Corresponding author; email: hawleyr{at}usa.redcross.org. 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'HS4 insulator 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: A. Ramezani and R. G. Hawley Correction of murine hemophilia A following nonmyeloablative transplantation of hematopoietic stem cells engineered to encode an enhanced human factor VIII variant using a safety-augmented retroviral vector Blood, July 16, 2009; 114(3): 526 - 534. [Abstract] [Full Text] [PDF] B. Y. Ryu, M. V. Evans-Galea, J. T. Gray, D. M. Bodine, D. A. Persons, and A. W. Nienhuis An experimental system for the evaluation of retroviral vector design to diminish the risk for proto-oncogene activation Blood, February 15, 2008; 111(4): 1866 - 1875. [Abstract] [Full Text] [PDF] J. He, Q. Yang, and L.-J. Chang Dynamic DNA Methylation and Histone Modifications Contribute to Lentiviral Transgene Silencing in Murine Embryonic Carcinoma Cells J. Virol., November 1, 2005; 79(21): 13497 - 13508. [Abstract] [Full Text] [PDF] P. Malik and P. I. Arumugam Gene Therapy for {beta}-Thalassemia Hematology, January 1, 2005; 2005(1): 45 - 50. [Abstract] [Full Text] [PDF] G. Puthenveetil, J. Scholes, D. Carbonell, N. Qureshi, P. Xia, L. Zeng, S. Li, Y. Yu, A. L Hiti, J.-K. Yee, et al. Successful correction of the human {beta}-thalassemia major phenotype using a lentiviral vector Blood, December 1, 2004; 104(12): 3445 - 3453. [Abstract] [Full Text] [PDF] P. Kurre, J. Morris, B. Thomasson, D. B. Kohn, and H.-P. Kiem Scaffold attachment region-containing retrovirus vectors improve long-term proviral expression after transplantation of GFP-modified CD34+ baboon repopulating cells Blood, November 1, 2003; 102(9): 3117 - 3119. [Abstract] [Full Text] [PDF] G. F. Atweh, J. DeSimone, Y. Saunthararajah, H. Fathallah, R. S. Weinberg, R. L. Nagel, M. E. Fabry, and R. J. Adams Hemoglobinopathies Hematology, January 1, 2003; 2003(1): 14 - 39. [Abstract] [Full Text] [PDF]

	Copyright © 2003 by American Society of Hematology         Online ISSN: 1528-0020