Okamoto Ikuhiro
Department of Anatomy and Cell Biology, Graduate School of Medicine, Kyoto University, Kyoto, Japan.
JST, ERATO, Kyoto, Japan.
Methods Mol Biol. 2018;1861:149-159. doi: 10.1007/978-1-4939-8766-5_12.
Transcriptional and epigenetic dynamics of the genome occur during early development in mammals. It has been difficult to study these dynamics due to the limitation of materials and the difficulty of handling. In this chapter, we describe our attempt to apply a combination of immunofluorescence (IF), and RNA and DNA fluorescent in situ hybridization (FISH) in preimplantation mouse embryos. We have concentrated on refining these techniques to study the dynamics of X chromosome inactivation in mouse embryos. The techniques and general underlying principles described here should be applicable to other mammals of interest.
基因组的转录和表观遗传动力学发生在哺乳动物的早期发育过程中。由于材料的限制和操作的难度,研究这些动力学一直很困难。在本章中,我们描述了我们尝试在植入前小鼠胚胎中应用免疫荧光(IF)以及RNA和DNA荧光原位杂交(FISH)相结合的方法。我们专注于完善这些技术,以研究小鼠胚胎中X染色体失活的动力学。这里描述的技术和一般基本原理应该适用于其他感兴趣的哺乳动物。