CAJ | 학술논문

piggyBac (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。
piggyBac (PB)전좌계통구유전좌효솔고、산제정학、반수궤삽입화휴대편단교대등우점。단시작위일충전기인실험적공구，특별시재포유동물개체수평적전기인방면，환수요제고기전기인효솔，병강저외원기인수궤삽입대내원기인파배적풍험。근년래적연구결과현시，PB 전좌계통득도료진일보개진：채용 PB 전좌매여DNA 특이성결합단백융합이구성적융합형전좌매，표현출외원편단유삽입도염색체파향위점적경향；채용돌변체사선적방법제고료PB전좌매적활성，획득료지구유절제활성이몰유삽입활성적신형PB전좌매；채용PB전좌계통여세균인공염색체(Bacterial artificial chromosomes, BAC)재체연합사휴대적외원편단장도제고도료207 kb。개진후적PB전좌계통재기인조연구、기인치료、유도다능간세포(Induced pluripotent stem cells, iPSCs)유도급기분화방면발휘료교대적작용。문장대PB전좌계통적최신연구진전화응용전경진행료종술。
ThepiggyBac(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 trans-posases obtained by mutagenesis have dramatically enhanced transposition activity and generated a novel function which is excision competent and integration defective. Furthermore, PB transposon system can carry large exogenous DNA fragments up to 207 kb when combining with the bacterial artificial chromosome vector. So far, these modified transposon systems have been widely applied in genome studies, gene therapy and induced pluripotent stem cells (iPS cells). In this study, we review the latest studies onpiggyBac transposon system and its application prospect.