Tanaka Satomi S, Yamaguchi Yasuka L, Jones Vanessa J, Tam Patrick P L
Department of Kidney Development, Institute of Molecular Embryology and Genetics, Kumamoto University, Kumamoto, Japan.
Methods Mol Biol. 2014;1092:367-93. doi: 10.1007/978-1-60327-292-6_22.
A well-established experimental paradigm to analyze gene function in development is to elucidate the impact of gain and loss of gene activity on cell differentiation, tissue modelling, organogenesis, and morphogenesis. This chapter describes the experimental protocols to study gene function by means of electroporation and lipofection to manipulate genetic activity in whole embryos and fetal organs in vitro. These techniques allow for more precise control of the timing, with reference to developmental age or stage, and the cell/tissue-specificity of the changes in gene activity. They provide an alternative strategy that can expedite the analysis of gene function before resorting to the conventional means of transgenesis and gene targeting in the whole organism.
一种用于分析发育过程中基因功能的成熟实验范式是阐明基因活性的增加和丧失对细胞分化、组织建模、器官发生和形态发生的影响。本章描述了通过电穿孔和脂质转染在体外操纵全胚胎和胎儿器官中的遗传活性来研究基因功能的实验方案。这些技术能够更精确地控制时间（参照发育年龄或阶段）以及基因活性变化的细胞/组织特异性。它们提供了一种替代策略，在诉诸全生物体中传统的转基因和基因靶向方法之前，可以加快基因功能的分析。
