Region-specific saturation germline mutagenesis in mice using the Sleeping Beauty transposon system

Vincent W. Keng; Kojiro Yae; Tomoko Hayakawa; Sumi Mizuno; Yoshihiro Uno; Kosuke Yusa; Chikara Kokubu; Taroh Kinoshita; Keiko Akagi; Nancy A. Jenkins; Neal G. Copeland; Kyoji Horie; Junji Takeda

doi:10.1038/nmeth795

Region-specific saturation germline mutagenesis in mice using the Sleeping Beauty transposon system

Vincent W. Keng, Kojiro Yae, Tomoko Hayakawa, Sumi Mizuno, Yoshihiro Uno, Kosuke Yusa, Chikara Kokubu, Taroh Kinoshita, Keiko Akagi, Nancy A. Jenkins, Neal G. Copeland, Kyoji Horie, Junji Takeda

Research output: Contribution to journal › Article › peer-review

101 Scopus citations

Abstract

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.

Original language	English (US)
Pages (from-to)	763-769
Number of pages	7
Journal	Nature Methods
Volume	2
Issue number	10
DOIs	https://doi.org/10.1038/nmeth795
State	Published - Oct 2005
Externally published	Yes

ASJC Scopus subject areas

Biotechnology
Biochemistry
Molecular Biology
Cell Biology

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title = "Region-specific saturation germline mutagenesis in mice using the Sleeping Beauty transposon system",

abstract = "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.",

author = "Keng, {Vincent W.} and Kojiro Yae and Tomoko Hayakawa and Sumi Mizuno and Yoshihiro Uno and Kosuke Yusa and Chikara Kokubu and Taroh Kinoshita and Keiko Akagi and Jenkins, {Nancy A.} and Copeland, {Neal G.} and Kyoji Horie and Junji Takeda",

note = "Funding Information: We thank G. Kondoh, M. Kouno, E.S. Saito and R. Ikeda for helpful advice and excellent technical assistance. We also thank S. Makino and Y. Odan for administrative assistance in preparing this manuscript. This work was supported by grants from the New Energy and Industrial Technology Development Organization of Japan; Uehara Memorial Foundation; Preventure Program, Japan Science and Technology Agency; RIKEN, The Institute of Physical and Chemical Research; and a grant-in-aid for Scientific Research from the Ministry of Education, Culture, Sports, Science and Technology of Japan.",

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language = "English (US)",

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AU - Keng, Vincent W.

AU - Yae, Kojiro

AU - Hayakawa, Tomoko

AU - Mizuno, Sumi

AU - Uno, Yoshihiro

AU - Yusa, Kosuke

AU - Kokubu, Chikara

AU - Kinoshita, Taroh

AU - Akagi, Keiko

AU - Jenkins, Nancy A.

AU - Copeland, Neal G.

AU - Horie, Kyoji

AU - Takeda, Junji

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Region-specific saturation germline mutagenesis in mice using the Sleeping Beauty transposon system

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