Skip Navigation

Nucleic Acids Research 2005 33(9):e85; doi:10.1093/nar/gni083
This Article
Right arrow Full Text Freely available
Right arrow Print PDF (896K) Freely available
Right arrow Screen PDF (266K) Freely available
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Email this article to a friend
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Add to My Personal Archive
Right arrow Download to citation manager
Right arrow Commercial Re-use Guidelines
for Open Access NAR Content
Google Scholar
Right arrow Articles by Kakitani, M.
Right arrow Articles by Tomizuka, K.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Kakitani, M.
Right arrow Articles by Tomizuka, K.
Related Collections
Right arrow Recombinant DNA expression
Right arrow DNA transfer
Social Bookmarking
Add to CiteULike   Add to Connotea   Add to Del.icio.us  
What's this?

Published online 24 May 2005

© The Author 2005. Published by Oxford University Press. All rights reserved
The online version of this article has been published under an open access model. Users are entitled to use, reproduce, disseminate, or display the open access version of this article for non-commercial purposes provided that: the original authorship is properly and fully attributed; the Journal and Oxford University Press are attributed as the original place of publication with the correct citation details given; if an article is subsequently reproduced or disseminated not in its entirety but only in part or as a derivative work this must be clearly indicated. For commercial re-use, please contact journals.permissions{at}oupjournals.org

Methods Online

A novel transgenic chimaeric mouse system for the rapid functional evaluation of genes encoding secreted proteins

Makoto Kakitani, Takeshi Oshima, Kaori Horikoshi, Tetsuo Yoshitome, Akiko Ueda, Miwa Kajikawa, Yumi Iba, Yoshinao Ozone, Yuki Ijima, Tohko Yoshino, Mikiko Itoh, Sachiko Seki, Ayako Aoki, Toshie Ishihara, Michiyo Shionoya, Utako Makino, Rina Kitada, Atsuko Ohguma, Takami Ohta, Yoshimasa Yoshida, Hiroe Kudoh1, Kazunori Hanaoka1, Kazunori Sibuya, Isao Ishida, Minoru Kakeda, Mikio Yagi, Takashi Yoneya and Kazuma Tomizuka*

Pharmaceutical Research Laboratory, Pharmaceutical Division, Kirin Brewery Co. Ltd. 3 Miyahara-cho, Takasaki-shi, Gunma 370-1295, Japan 1Molecular Embryology, Department of Biosciences, School of Science, Kitasato University 1-15-1 Kitasato, Sagamihara-shi, Kanagawa, 228-8555, Japan

*To whom correspondence should be addressed. Tel: +81 27 346 9934; Fax: +81 27 346 1971; Email: ktomizuka{at}kirin.co.jp

Received February 12, 2005. Revised May 2, 2005. Accepted May 2, 2005.

A major challenge of the post-genomic era is the functional characterization of anonymous open reading frames (ORFs) identified by the Human Genome Project. In this context, there is a strong requirement for the development of technologies that enhance our ability to analyze gene functions at the level of the whole organism. Here, we describe a rapid and efficient procedure to generate transgenic chimaeric mice that continuously secrete a foreign protein into the systemic circulation. The transgene units were inserted into the genomic site adjacent to the endogenous immunoglobulin (Ig) {kappa} locus by homologous recombination, using a modified mouse embryonic stem (ES) cell line that exhibits a high frequency of homologous recombination at the Ig{kappa} region. The resultant ES clones were injected into embryos derived from a B-cell-deficient host strain, thus producing chimaerism-independent, B-cell-specific transgene expression. This feature of the system eliminates the time-consuming breeding typically implemented in standard transgenic strategies and allows for evaluating the effect of ectopic transgene expression directly in the resulting chimaeric mice. To demonstrate the utility of this system we showed high-level protein expression in the sera and severe phenotypes in human EPO (hEPO) and murine thrombopoietin (mTPO) transgenic chimaeras.


Add to CiteULike CiteULike   Add to Connotea Connotea   Add to Del.icio.us Del.icio.us    What's this?


This article has been cited by other articles:


Home page
 ScienceHome page
K.-A. Kim, M. Kakitani, J. Zhao, T. Oshima, T. Tang, M. Binnerts, Y. Liu, B. Boyle, E. Park, P. Emtage, et al.
Mitogenic Influence of Human R-Spondin1 on the Intestinal Epithelium
Science, August 19, 2005; 309(5738): 1256 - 1259.
[Abstract] [Full Text] [PDF]


Disclaimer: Please note that abstracts for content published before 1996 were created through digital scanning and may therefore not exactly replicate the text of the original print issues. All efforts have been made to ensure accuracy, but the Publisher will not be held responsible for any remaining inaccuracies. If you require any further clarification, please contact our Customer Services Department.