Suganuma Ryota, Pelczar Pawel, Spetz Jean François, Hohn Barbara, Yanagimachi Ryuzo, Moisyadi Stefan
Department of Anatomy and Reproductive Biology, University of Hawaii School of Medicine, Honolulu, Hawaii 96822, USA.
Biol Reprod. 2005 Dec;73(6):1157-63. doi: 10.1095/biolreprod.105.044669. Epub 2005 Aug 3.
We have developed a novel method for mouse transgenesis. The procedure relies on a hyperactive Tn5 transposase to insert a transgene into mouse chromosomes during intracytoplasmic sperm injection. This procedure integrates foreign DNA into the mouse genome with dramatically increased effectiveness as compared to conventional methods such as pronuclear microinjection and traditional sperm injection-mediated transgenesis. Our data indicate that with this method, transgenic mice, both hybrids and inbreds, can be produced more consistently and with lower numbers of manipulated oocytes required for traditional microinjection methods. The transposase-mediated transgenesis technique is also effective with round spermatids, offering the potential for rescuing the fertility of azoospermic animals using sperm precursor cells.
我们开发了一种用于小鼠转基因的新方法。该程序依赖于一种超活性Tn5转座酶，在胞浆内精子注射过程中将转基因插入小鼠染色体。与传统方法如原核显微注射和传统精子注射介导的转基因方法相比，该程序将外源DNA整合到小鼠基因组中的效率显著提高。我们的数据表明，使用这种方法，可以更稳定地产生转基因小鼠，包括杂种和近交系小鼠，并且与传统显微注射方法相比，所需操作的卵母细胞数量更少。转座酶介导的转基因技术对圆形精子细胞也有效，这为使用精子前体细胞挽救无精子症动物的生育能力提供了潜力。
