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Prepublished online as a Blood First Edition Paper on July 24, 2003; DOI 10.1182/blood-2003-05-1424. This Article 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 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. Social Bookmarking                   What's this? Submitted May 6, 2003 Accepted July 15, 2003 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 Kuhlcke, Birgit Stahle, Axel R Zander, Christopher Baum, and Boris Fehse* Department of Bone Marrow Transplantation, University Hospital Hamburg-Eppendorf, Hamburg, Germany EUFETS AG, Idar-Oberstein, Germany Department of Hematology and Oncology, Hannover Medical School, Hannover, Germany Division of Experimental Hematology, Cincinnati Children's Research Foundation, Cincinnati, OH, USA * Corresponding author; email: fehse{at}uke.uni-hamburg.de. Retroviral vectors are commonly used in clinical gene therapy, but recent observations of insertional oncogene activation in preclinical and clinical settings have enforced the discussion on 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 both K562 leukemia and primary CD34+ cells. We found an exponential increase of integration numbers in correlation to gene transfer rates, and a linear increase of expression levels with insertion frequency. In average we detected one vector insertion per transduced cell for a gene transfer of <30%, for 60% three and for 90% (in K562) about nine. Clonal analysis revealed strikingly increased variations of both transgene copy numbers (>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 generation of clones containing multiple insertions. CiteULike    Connotea    Del.icio.us    Digg    Reddit    Technorati    What's this? This article has been cited by other articles: S. Okamoto, J. Mineno, H. 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