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Blood, 15 August 2005, Vol. 106, No. 4, pp. 1147-1153. Prepublished online as a Blood First Edition Paper on May 10, 2005; DOI 10.1182/blood-2004-12-4839. This Article Full Text (PDF) All Versions of this Article: 2004-12-4839v1 106/4/1147    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 Bot, I. Articles by Biessen, E. A Search for Related Content PubMed PubMed Citation Articles by Bot, I. Articles by Biessen, E. A Related Collections Related Article in Blood Online Social Bookmarking                   What's this?  Previous Article  |  Next Article  Submitted December 21, 2004 Accepted April 22, 2005 Lentiviral shRNA silencing of murine bone marrow cell CCR2 leads to persistent knockdown of CCR2 function in vivo Ilze Bot*, Jian Guo, Miranda Van Eck, Peter J Van Santbrink, Pieter H Groot, Reeni B Hildebrand, Jurgen Seppen, Theo J Van Berkel, and Erik A Biessen Division of Biopharmaceutics, Gorlaeus Laboratories, Leiden University, Leiden, The Netherlands Department of Experimental Hepatology, Academic Medical Center, Amsterdam, The Netherlands * Corresponding author; email: i.bot{at}lacdr.leidenuniv.nl. A major barrier in hematopoietic gene function studies is posed by the laborious and time-consuming generation of knockout mice with an appropriate genetic background. Here we present a novel lentivirus based strategy for the in situ generation of hematopoietic knock-downs. A short hairpin RNA (shRNA) was designed targeting murine CC-chemokine receptor 2 (CCR2), which was able to specifically blunt CCR2 expression at mRNA, protein and functional level in vitro. Reconstitution of irradiated recipient mice with autologous bone marrow, that had been ex vivo transduced with shRNA lentivirus, led to persistent down-regulation of CCR2 expression, which translated into a 70% reduction in CCR2 dependent recruitment of macrophages to an inflamed peritoneal cavity without noticeable side-effects on related chemokine receptors or general inflammation status. These findings clearly demonstrate the potential of shRNA lentivirus infected bone marrow transplantation as a rapid and effective method to generate hematopoietic knockdowns for leukocyte gene function studies. CiteULike    Connotea    Del.icio.us    Digg    Reddit    Technorati    What's this? Related Article in Blood Online: Hematopoiesis: yet another milestone for RNAi Mikko O. Laukkanen Blood 2005 106: 1143. [Full Text] [PDF] This article has been cited by other articles: C. C. Porter and J. DeGregori Interfering RNA-mediated purine analog resistance for in vitro and in vivo cell selection Blood, December 1, 2008; 112(12): 4466 - 4474. [Abstract] [Full Text] [PDF]

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