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Blood, 15 March 2005, Vol. 105, No. 6, pp. 2307-2315. Prepublished online as a Blood First Edition Paper on November 12, 2004; DOI 10.1182/blood-2004-03-0798. This Article Full Text Full Text (PDF) All Versions of this Article: 2004-03-0798v1 105/6/2307    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 De Palma, M. Articles by Naldini, L. Search for Related Content PubMed PubMed Citation Articles by De Palma, M. Articles by Naldini, L. Related Collections Hematopoiesis and Stem Cells Gene Therapy Social Bookmarking                   What's this?  Previous Article  |  Table of Contents  |  Next Article  GENE THERAPY Promoter trapping reveals significant differences in integration site selection between MLV and HIV vectors in primary hematopoietic cells Michele De Palma, Eugenio Montini, Francesca R. Santoni de Sio, Fabrizio Benedicenti, Alessandra Gentile, Enzo Medico, and Luigi Naldini From the H. San Raffaele-Telethon Institute for Gene Therapy (HSR-TIGET), Milan, Italy; the Vita Salute San Raffaele University, H. San Raffaele Scientific Institute, Milan, Italy; and the Institute for Cancer Research and Treatment (IRCC), Torino, Italy. Recent reports have indicated that human immunodeficiency virus (HIV) and murine leukemia virus (MLV) vectors preferentially integrate into active genes. Here, we used a novel approach based on genetic trapping to rapidly score several thousand integration sites and found that MLV vectors trapped cellular promoters more efficiently than HIV vectors. Remarkably, 1 in 5 MLV integrations trapped an active promoter in different cell lines and primary hematopoietic cells. Such frequency was even higher in growth-stimulated lymphocytes. We show that the different behavior of MLV and HIV vectors was dependent on a different integration pattern within transcribed genes. Whereas MLV-based traps showed a strong bias for promoter-proximal integration leading to efficient reporter expression, HIV-based traps integrated throughout transcriptional units and were limited for expression by the distance from the promoter and the reading frame of the targeted gene. Our results indicate a strong propensity of MLV to establish transcriptional interactions with cellular promoters, a behavior that may have evolved to enhance proviral expression and may increase the insertional mutagenesis risk. Promoter trapping efficiency provides a convenient readout to assess transcriptional interactions between the vector and its flanking genes at the integration site and to compare integration site selection among different cell types and in different growth conditions. CiteULike    Connotea    Del.icio.us    Digg    Reddit    Technorati    What's this? This article has been cited by other articles: G. P. Wang, A. Garrigue, A. Ciuffi, K. Ronen, J. Leipzig, C. Berry, C. Lagresle-Peyrou, F. Benjelloun, S. Hacein-Bey-Abina, A. Fischer, et al. DNA bar coding and pyrosequencing to analyze adverse events in therapeutic gene transfer Nucleic Acids Res., May 1, 2008; 36(9): e49 - e49. [Abstract] [Full Text] [PDF] L. Gammaitoni, S. Lucchi, S. Bruno, M. Tesio, M. Gunetti, Y. Pignochino, G. Migliardi, L. Lazzari, M. Aglietta, P. Rebulla, et al. Serial Transplantations in Nonobese Diabetic/Severe Combined Immunodeficiency Mice of Transduced Human CD34+ Cord Blood Cells: Efficient Oncoretroviral Gene Transfer and Ex Vivo Expansion Under Serum-Free Conditions Stem Cells, May 1, 2006; 24(5): 1201 - 1212. [Abstract] [Full Text] [PDF]

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