piggyBac转座系统的功能改进及其在哺乳动物中的应用.pdfVIP

Hereditas (Beijing) 2014 年 10 月, 36(10): 965 ―973 综 述 piggy Bac 转座系统的功能改进及在哺乳动物中的应用 钱秋杰，车家倩，叶露鹏，钟伯雄 浙江大学动物科学学院，杭州 310058 摘要:piggy Bac (PB)转座系统具有转座效率高、删除精确、半随机插入和携带片段较大等优点。但是作为一种 转基因实验的工具，特别是在哺乳动物个体水平的转基因方面，还需要提高其转基因效率，并降低外源基因随 机插入对内源基因破坏的风险。近年来的研究结果显示，PB 转座系统得到了进一步改进：采用 PB 转座酶与 DNA 特异性结合蛋白融合而构成的融合型转座酶，表现出外源片段有插入到染色体靶向位点的倾向；采用突 变体筛选的方法提高了PB 转座酶的活性，获得了只具有切除活性而没有插入活性的新型PB 转座酶；采用PB 转座系统与细菌人工染色体(Bacterial artificial chromosomes, BAC)载体联合使携带的外源片段长度提高到了 207 kb 。改进后的PB 转座系统在基因组研究、基因治疗、诱导多能干细胞(Induced pluripotent stem cells, iPSCs) 诱导及其分化方面发挥了较大的作用。文章对PB 转座系统的最新研究进展和应用前景进行了综述。 关键词: piggy Bac ；转基因；靶向插入；转座 The improvement and application of piggy Bac transposon system in mammals Qiujie Qian, Jiaqian Che, Lupeng Ye, Boxiong Zhong College of Animal Science, Zhejiang University, Hangzhou 310058, China Abstract: The piggy Bac (PB) transposon system is a useful genomic engineering tool due to its high transposition efficiency, precise excision, semi-random insertion and large cargo capacity. But, it still needs to further improve the transgenic efficiency and reduce the risk of endogenous disruption caused by the random insertion of exogenous gene, especially in transgenic experiments of individual mammals. In recent studies, the PB transposase is fused with a DNA binding protein as a chimeric protein, which can guide the transposon to pre-designed loci. Besides, PB transposases obtained by mutagenesis have dramatically enhanced transposition activity and generated a novel func- tion which is excision competent and integration defective. Furthermore, PB transposon system can carry large ex- ogenous DNA fragments up to 207 kb when combining with the bacterial artificial chromosome vector. So far, these modified tra