Combined Immunofluorescence, RNA FISH, and DNA FISH in Preimplantation Mouse Embryos
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.
Key wordsImmunofluorescence RNA FISH DNA FISH Preimplantation mouse embryos X-chromosome inactivation
We thank E. Heard for critical reading of the manuscript.
This work was supported by funds from MEXT and JST-ERATO and by JSPS KAKENHI Grant Number JP25291076 to I.O.
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