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Nucleic Acids Research 2004 32(13):e109; doi:10.1093/nar/gnh093
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Published online 22 July 2004

Nucleic Acids Research, Vol. 32 No. 13 © Oxford University Press 2004; all rights reserved

Atelocollagen-mediated synthetic small interfering RNA delivery for effective gene silencing in vitro and in vivo

Yoshiko Minakuchi, Fumitaka Takeshita1, Nobuyoshi Kosaka2, Hideo Sasaki1, Yusuke Yamamoto2, Makiko Kouno3, Kimi Honma3, Shunji Nagahara, Koji Hanai, Akihiko Sano, Takashi Kato2, Masaaki Terada1 and Takahiro Ochiya1,*

Formulation Research Laboratories, Sumitomo Pharmaceuticals Co. Ltd, Osaka 567-0878, Japan, 1 National Cancer Center Research Institute, 1-1, Tsukiji 5-chome, Chuo-ku, Tokyo 104-0045, Japan, 2 Department of Biology, School of Education, Waseda University, Tokyo 169-0051, Japan and 3 Koken Bioscience Institute, Tokyo 115-0051, Japan

* To whom correspondence should be addressed. Tel: +81 3 3542 2511; Fax: +81 3 3541 2685; Email: tochiya{at}ncc.go.jp

Received April 8, 2004; Revised May 26, 2004; Accepted June 7, 2004

Silencing gene expression by siRNAs is rapidly becoming a powerful tool for the genetic analysis of mammalian cells. However, the rapid degradation of siRNA and the limited duration of its action call for an efficient delivery technology. Accordingly, we describe here that Atelocollagen complexed with siRNA is resistant to nucleases and is efficiently transduced into cells, thereby allowing long-term gene silencing. Site-specific in vivo administration of an anti-luciferase siRNA/Atelocollagen complex reduced luciferase expression in a xenografted tumor. Furthermore, Atelocollagen-mediated transfer of siRNA in vivo showed efficient inhibition of tumor growth in an orthotopic xenograft model of a human non-seminomatous germ cell tumor. Thus, for clinical applications of siRNA, an Atelocollagen-based non-viral delivery method could be a reliable approach to achieve maximal function of siRNA in vivo.


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