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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.

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PLENARY PAPERS

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. E. Groot, Reeni B. Hildebrand, Jurgen Seppen, Theo J. C. Van Berkel, and Erik A. L. Biessen

From the Division of Biopharmaceutics, Gorlaeus Laboratories, Leiden University, Leiden, The Netherlands; and the Academic Medical Center, Department of Experimental Hepatology, Amsterdam, The Netherlands.

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 knockdowns. A short hairpin RNA (shRNA) was designed targeting murine CC-chemokine receptor 2 (CCR2), which was able to specifically blunt CCR2 expression at the mRNA, protein, and functional levels 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.


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Blood 2005 106: 1143. [Full Text] [PDF]



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