Center for Development of Advanced Medical Technology, Jichi Medical University, Tochigi, Japan.
Faculty of Bioscience and Bioindustry, Tokushima University, Tokushima, Japan.
Methods Mol Biol. 2023;2637:293-300. doi: 10.1007/978-1-0716-3016-7_22.
Pigs are excellent large animal models owing to their several physiological and anatomical similarities to humans. Somatic cell nuclear transfer using gene-modified cells is the mainstream approach for generating genetically modified pigs. Recent advances in improving gene editors such as the CRISPR/Cas9 system have enabled direct gene modification in zygotes/embryos. Here, we describe the gene editing by electroporation of Cas9 protein (GEEP) method, an optimized electroporation-mediated method for the introduction of CRISPR/Cas9 into porcine zygotes/embryos. The simplicity and micromanipulation-free procedures are the major advantages of this method.
猪由于其与人类在生理和解剖学上的许多相似之处，是优秀的大型动物模型。使用基因修饰细胞的体细胞核移植是生成基因修饰猪的主流方法。近年来，基因编辑技术如 CRISPR/Cas9 系统的改进，使得在受精卵/胚胎中直接进行基因修饰成为可能。在这里，我们描述了 Cas9 蛋白电穿孔（GEEP）法的基因编辑，这是一种优化的电穿孔介导的将 CRISPR/Cas9 导入猪受精卵/胚胎的方法。该方法的主要优点是简单且无需进行显微操作。
