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Molecular and Cellular Biology, December 2003, p. 9189-9207, Vol. 23, No. 24
0270-7306/03/$08.00+0     DOI: 10.1128/MCB.23.24.9189-9207.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Characterization of Sleeping Beauty Transposition and Its Application to Genetic Screening in Mice

Kyoji Horie,1,2 Kosuke Yusa,2 Kojiro Yae,2,3 Junko Odajima,4 Sylvia E. J. Fischer,5 Vincent W. Keng,2,6 Tomoko Hayakawa,2 Sumi Mizuno,2,6 Gen Kondoh,2 Takashi Ijiri,7 Yoichi Matsuda,7,8 Ronald H. A. Plasterk,5 and Junji Takeda1,2,6*

Collaborative Research Center for Advanced Science and Technology,1 Department of Social and Environmental Medicine,2 Department of Hematology and Oncology,4 Japan Science and Technology Corporation, Graduate School of Medicine, Osaka University, Suita, Osaka 565-0871,6 Division of Gene Function in Animals, Nara Institute of Science and Technology, Ikoma, Nara 630-0101,3 Laboratory of Cytogenetics, Division of Bioscience, Graduate School of Environmental Earth Science,7 Laboratory of Animal Cytogenetics, Center for Advanced Science and Technology, Hokkaido University, Kita-ku, Sapporo 060-0810, Japan,8 Hubrecht Laboratory, Center for Biomedical Genetics, 3584 CT Utrecht, The Netherlands5

Received 7 July 2003/ Returned for modification 22 August 2003/ Accepted 17 September 2003

The use of mutant mice plays a pivotal role in determining the function of genes, and the recently reported germ line transposition of the Sleeping Beauty (SB) transposon would provide a novel system to facilitate this approach. In this study, we characterized SB transposition in the mouse germ line and assessed its potential for generating mutant mice. Transposition sites not only were clustered within 3 Mb near the donor site but also were widely distributed outside this cluster, indicating that the SB transposon can be utilized for both region-specific and genome-wide mutagenesis. The complexity of transposition sites in the germ line was high enough for large-scale generation of mutant mice. Based on these initial results, we conducted germ line mutagenesis by using a gene trap scheme, and the use of a green fluorescent protein reporter made it possible to select for mutant mice rapidly and noninvasively. Interestingly, mice with mutations in the same gene, each with a different insertion site, were obtained by local transposition events, demonstrating the feasibility of the SB transposon system for region-specific mutagenesis. Our results indicate that the SB transposon system has unique features that complement other mutagenesis approaches.

* Corresponding author. Mailing address: Department of Social and Environmental Medicine H3, Graduate School of Medicine, Osaka University, 2-2 Yamadaoka, Suita, Osaka 565-0871, Japan. Phone: 81 6 6879 3262. Fax: 81 6 6879 3266. E-mail: takeda{at}mr-envi.med.osaka-u.ac.jp.

Molecular and Cellular Biology, December 2003, p. 9189-9207, Vol. 23, No. 24
0022-538X/03/$08.00+0     DOI: 10.1128/MCB.23.24.9189-9207.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



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