Region-specific saturation germline mutagenesis in mice using the Sleeping Beauty transposon system
Region-specific saturation germline mutagenesis in mice using the Sleeping Beauty transposon system
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DOI：
10.1038/nmeth795
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发表时间：
2005-10-01
期刊：
影响因子：
48
通讯作者：
Takeda, J
中科院分区：
文献类型：
--
作者：
Keng, VW;Yae, K;Takeda, J
最近整合全基因组序列与全基因组转录组获利揭示了基因组内特定染色体区域的功能单位的存在，如在基因簇的协调表达和功能相关基因的共定位中所见。有效的区域特异性突变筛选将极大地促进解决这些簇重要性的研究。在这里，我们使用的“本地跳跃”的DNA型转座子，睡美人（SB）的现象，区域特异性饱和诱变。一种含有转座子（作为诱变剂）和转座酶（识别和动员转座子）的转基因小鼠被培育用于生殖细胞转座事件，使我们能够产生许多突变小鼠。原始供体位点的4-Mb区域内的所有基因被SB突变，表明该系统在特定染色体区域内用于功能基因组研究的潜力。
Recent consolidation of the whole-genome sequence with genome-wide transcriptome profiting revealed the existence of functional units within the genome in specific chromosomal regions, as seen in the coordinated expression of gene clusters and colocatization 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.