Department of Anatomy, John A. Burns School of Medicine, University of Hawaii, Honolulu, HI 96822, USA.
Proc Natl Acad Sci U S A. 2012 Nov 20;109(47):19184-9. doi: 10.1073/pnas.1216473109. Epub 2012 Oct 23.
We have developed a unique method for mouse transgenesis. The transposase-enhanced pronuclear microinjection (PNI) technique described herein uses the hyperactive piggyBac transposase to insert a large transgene into the mouse genome. This procedure increased transgene integration efficiency by fivefold compared with conventional PNI or intracytoplasmic sperm injection-mediated transgenesis. Our data indicate that the transposase-enhanced PNI technique additionally requires fewer embryos to be microinjected than traditional methods to obtain transgenic animals. This transposase-mediated approach is also very efficient for single-cell embryo cytoplasmic injections, offering an easy-to-implement transgenesis method to the scientific community.
我们开发了一种独特的小鼠转基因方法。本文描述的转座酶增强的原核显微注射（PNI）技术使用超活性 piggyBac 转座酶将一个大的转基因插入小鼠基因组中。与传统的 PNI 或胞质内精子注射介导的转基因相比，该程序将转基因整合效率提高了五倍。我们的数据表明，与传统方法相比，转座酶增强的 PNI 技术需要更少的胚胎进行显微注射，以获得转基因动物。这种转座酶介导的方法对于单细胞胚胎细胞质注射也非常有效，为科学界提供了一种易于实施的转基因方法。
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