Nature Methods
2, 763 - 769 (2005)
Published online: 22 September 2005; | doi:10.1038/nmeth795
Region-specific saturation germline mutagenesis in mice using the Sleeping Beauty transposon systemVincent W Keng1, Kojiro Yae2, Tomoko Hayakawa2, Sumi Mizuno1, Yoshihiro Uno3, Kosuke Yusa2, Chikara Kokubu1, 2, Taroh Kinoshita4, Keiko Akagi5, Nancy A Jenkins5, Neal G Copeland5, Kyoji Horie2
& Junji Takeda1, 21
Center for Advanced Science and Innovation, Osaka University, 2-1 Yamadaoka, Suita, Osaka 565-0871, Japan. 2
Department of Social and Environmental Medicine, Graduate School of Medicine, Osaka University, 2-2 Yamadaoka, Suita, Osaka 565-0871, Japan. 3
Experimental Animal Science, Graduate School of Medicine, Osaka University, 2-2 Yamadaoka, Suita, Osaka 565-0871, Japan. 4
Research Institute for Microbial Diseases, Osaka University, 3-1 Yamadaoka, Suita, Osaka 565-0871, Japan. 5
Mouse Cancer Genetics Program, National Cancer Institute, Frederick, Maryland 21701, USA.
Correspondence should be addressed to Junji Takeda takeda@mr-envi.med.osaka-u.ac.jp Recent consolidation of the whole-genome sequence with genome-wide transcriptome profiling revealed the existence of functional units within the genome in specific chromosomal regions, as seen in the coordinated expression of gene clusters and colocalization of functionally related genes. An efficient region-specific mutagenesis screen would greatly facilitate research in addressing the importance of these clusters. Here we use the 'local hopping' phenomenon of a DNA-type transposon, Sleeping Beauty (SB), for region-specific saturation mutagenesis. A transgenic mouse containing both transposon (acts as a mutagen) and transposase (recognizes and mobilizes the transposon) was bred for germ-cell transposition events, allowing us to generate many mutant mice. All genes within a 4-Mb region of the original donor site were mutated by SB, indicating the potential of this system for functional genomic studies within a specific chromosomal region.
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