Horie Kyoji, Yusa Kosuke, Yae Kojiro, Odajima Junko, Fischer Sylvia E J, Keng Vincent W, Hayakawa Tomoko, Mizuno Sumi, Kondoh Gen, Ijiri Takashi, Matsuda Yoichi, Plasterk Ronald H A, Takeda Junji
Collaborative Research Center for Advanced Science and Technology, Department of Social and Environmental Medicine, Graduate School of Medicine, Osaka University, 2-2 Yamadaoka, Suita, Osaka 565-0871, Japan.
Mol Cell Biol. 2003 Dec;23(24):9189-207. doi: 10.1128/MCB.23.24.9189-9207.2003.
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.
突变小鼠的使用在确定基因功能方面起着关键作用,最近报道的睡美人(SB)转座子的种系转座将提供一个新的系统来促进这种方法。在本研究中,我们对小鼠种系中的SB转座进行了表征,并评估了其产生突变小鼠的潜力。转座位点不仅聚集在供体位点附近的3 Mb范围内,而且在该簇之外广泛分布,这表明SB转座子可用于区域特异性诱变和全基因组诱变。种系中转座位点的复杂性足以大规模产生突变小鼠。基于这些初步结果,我们使用基因捕获方案进行种系诱变,并且使用绿色荧光蛋白报告基因使得能够快速、非侵入性地选择突变小鼠。有趣的是,通过局部转座事件获得了同一基因发生突变但插入位点不同的小鼠,证明了SB转座子系统用于区域特异性诱变的可行性。我们的结果表明,SB转座子系统具有独特的特征,可补充其他诱变方法。