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Blood, 1 December 2003, Vol. 102, No. 12, pp. 3934-3937. Prepublished online as a Blood First Edition Paper on July 24, 2003; DOI 10.1182/blood-2003-05-1424. This Article Full Text Full Text (PDF) All Versions of this Article: 2003-05-1424v1 102/12/3934    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 Kustikova, O. S. Articles by Fehse, B. Search for Related Content PubMed PubMed Citation Articles by Kustikova, O. S. Articles by Fehse, B. Related Collections Neoplasia Oncogenes and Tumor Suppressors Brief Reports Gene Therapy Social Bookmarking                   What's this?  Previous Article  |  Table of Contents  |  Next Article  GENE THERAPY Brief report Dose finding with retroviral vectors: correlation of retroviral vector copy numbers in single cells with gene transfer efficiency in a cell population Olga S. Kustikova, Anke Wahlers, Klaus Kühlcke, Birgit Stähle, Axel R. Zander, Christopher Baum, and Boris Fehse From the Department of Bone Marrow Transplantation, University Hospital Eppendorf, Hamburg, Germany; Europäisches Institut für Forschung und Entwicklung von Transplantationsstrategien (EUFETS) AG, Idar-Oberstein, Germany; Department of Hematology and Oncology, Hannover Medical School, Germany; and Division of Experimental Hematology, Cincinnati Children's Research Foundation, OH. Retroviral vectors are commonly used in clinical gene therapy, but recent observations of insertional oncogene activation in preclinical and clinical settings have forced a discussion of their safety. Here we investigated the relationship between retroviral transduction efficiency in mass cultures and the actual number of integrated vector copies in single cells using K562 leukemia and primary CD34+ cells. We found an exponential increase of integration numbers correlated to gene transfer rates and a linear increase of expression levels with insertion frequency. On average we detected one vector insertion per transduced cell for a gene transfer of less than 30%, 3 for 60%, and approximately 9 for 90% (in K562). Clonal analysis revealed strikingly increased variations of both transgene copy numbers (more than 20-fold in primary cells) and expression levels associated with higher transduction. Therefore, limiting retroviral gene transfer to approximately 30% may be suggested to avoid generating clones containing multiple insertions. CiteULike    Connotea    Del.icio.us    Digg    Reddit    Technorati    What's this? This article has been cited by other articles: Y.-J. Kim, Y.-S. Kim, A. Larochelle, G. Renaud, T. G. Wolfsberg, R. Adler, R. E. Donahue, P. Hematti, B.-K. Hong, J. Roayaei, et al. Sustained high-level polyclonal hematopoietic marking and transgene expression 4 years after autologous transplantation of rhesus macaques with SIV lentiviral vector-transduced CD34+ cells Blood, May 28, 2009; 113(22): 5434 - 5443. [Abstract] [Full Text] [PDF] A. M. Mahale, Z. A.T. Khan, M. Igarashi, G. J. Nanjangud, R. F. Qiao, S. Yao, S. W. Lee, and S. A. Aaronson Clonal Selection in Malignant Transformation of Human Fibroblasts Transduced with Defined Cellular Oncogenes Cancer Res., March 1, 2008; 68(5): 1417 - 1426. [Abstract] [Full Text] [PDF] S. Burghoff, Z. Ding, S. Godecke, A. 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