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Blood, 15 October 2006, Vol. 108, No. 8, pp. 2545-2553. Prepublished online as a Blood First Edition Paper on July 6, 2006; DOI 10.1182/blood-2005-08-024976. This Article Full Text Full Text (PDF) Supplemental Table and Figure All Versions of this Article: blood-2005-08-024976v1 108/8/2545    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 Modlich, U. Articles by Baum, C. Search for Related Content PubMed PubMed Citation Articles by Modlich, U. Articles by Baum, C. Related Collections Gene Therapy Related Article in Blood Online Social Bookmarking                   What's this?  Previous Article  |  Table of Contents  |  Next Article  GENE THERAPY Cell-culture assays reveal the importance of retroviral vector design for insertional genotoxicity Ute Modlich, Jens Bohne, Manfred Schmidt, Christof von Kalle, Sabine Knöss, Axel Schambach, and Christopher Baum From the Department of Experimental Hematology, Hannover Medical School, Germany; the Department of Hematology/Oncology and Institute for Molecular Medicine and Cell Research, Albert Ludwig University, Freiburg, Germany; the National Center for Tumor Diseases (NCT), Heidelberg, Germany; and the Division of Experimental Hematology, Cincinnati Children's Hospital Medical Center, OH. Retroviral vectors with long terminal repeats (LTRs), which contain strong enhancer/promoter sequences at both ends of their genome, are widely used for stable gene transfer into hematopoietic cells. However, recent clinical data and mouse models point to insertional activation of cellular proto-oncogenes as a dose-limiting side effect of retroviral gene delivery that potentially induces leukemia. Self-inactivating (SIN) retroviral vectors do not contain the terminal repetition of the enhancer/promoter, theoretically attenuating the interaction with neighboring cellular genes. With a new assay based on in vitro expansion of primary murine hematopoietic cells and selection in limiting dilution, we showed that SIN vectors using a strong internal retroviral enhancer/promoter may also transform cells by insertional mutagenesis. Most transformed clones, including those obtained after dose escalation of SIN vectors, showed insertions upstream of the third exon of Evi1 and in reverse orientation to its transcriptional orientation. Normalizing for the vector copy number, we found the transforming capacity of SIN vectors to be significantly reduced when compared with corresponding LTR vectors. Additional modifications of SIN vectors may further increase safety. Improved cell-culture assays will likely play an important role in the evaluation of insertional mutagenesis. CiteULike    Connotea    Del.icio.us    Digg    Reddit    Technorati    What's this? Related Article in Blood Online: The SINister side of retrovirus vectors David W. Emery Blood 2006 108: 2495-2496. [Full Text] [PDF] 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] Y.-J. Kim, Y.-S. Kim, A. Larochelle, G. Renaud, T. G. Wolfsberg, R. Adler, R. E. 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