Ding Sheng, Wu Xiaohui, Li Gang, Han Min, Zhuang Yuan, Xu Tian
Institute of Developmental Biology and Molecular Medicine, School of Life Sciences, Fudan University, 220 Handan Road, Shanghai 200433, China.
Cell. 2005 Aug 12;122(3):473-83. doi: 10.1016/j.cell.2005.07.013.
Transposable elements have been routinely used for genetic manipulation in lower organisms, including generating transgenic animals and insertional mutagenesis. In contrast, the usage of transposons in mice and other vertebrate systems is still limited due to the lack of an efficient transposition system. We have tested the ability of piggyBac (PB), a DNA transposon from the cabbage looper moth Trichoplusia ni, to transpose in mammalian systems. We show that PB elements carrying multiple genes can efficiently transpose in human and mouse cell lines and also in mice. PB permits the expression of the marker genes it carried. During germline transposition, PB could excise precisely from original insertion sites and transpose into the mouse genome at diverse locations, preferably transcription units. These data provide a first and critical step toward a highly efficient transposon system for a variety of genetic manipulations including transgenesis and insertional mutagenesis in mice and other vertebrates.
转座元件已被常规用于低等生物的基因操作，包括生成转基因动物和插入诱变。相比之下，由于缺乏高效的转座系统，转座子在小鼠和其他脊椎动物系统中的应用仍然有限。我们测试了来自甘蓝夜蛾Trichoplusia ni的DNA转座子piggyBac（PB）在哺乳动物系统中转座的能力。我们表明，携带多个基因的PB元件能够在人源和小鼠细胞系以及小鼠体内高效转座。PB允许其所携带的标记基因表达。在种系转座过程中，PB能够精确地从原始插入位点切除，并转座到小鼠基因组的不同位置，优先转座到转录单元。这些数据为构建一个高效转座子系统迈出了关键的第一步，该系统可用于包括小鼠和其他脊椎动物的转基因和插入诱变在内的各种基因操作。
