title: Analysis of nuclear reprogramming following nuclear transfer to Xenopus oocyte.
abstract: Germinal vesicle of stage V-VI Xenopus Laevis oocytes (at the prophase I stage of meiosis) can be used to transplant mammalian nuclei. In this type of interspecies nuclear transfer no cell division occurs and no new cell types are generated. However, the transplanted nuclei undergo extensive transcriptional reprogramming. Here, it is first explained how to carry out transplantation of multiple mammalian cell nuclei to Xenopus oocytes. It is then described how to perform RT-qPCR, Western Blot, Chromatin Immunoprecipitation, and live imaging analysis to monitor transcriptional reprogramming of the nuclei transplanted to oocytes.
authors.affiliation: Wellcome Trust/Cancer Research UK Gurdon Institute, The Henry Wellcome Building of Cancer and Developmental Biology, University of Cambridge, Tennis Court Road, Cambridge, CB2 1QN, UK, jj256@cam.ac.uk.
authors.forename: Jerome
authors.initials: J
authors.lastname: Jullien
mesh_terms: D000818:Animals; D015153:Blotting, Western; D002467:Cell Nucleus; D065150:Cellular Reprogramming; D047369:Chromatin Immunoprecipitation; D005260:Female; D007091:Image Processing, Computer-Assisted / Q000295:instrumentation / Q000379:methods*; D008845:Microinjections / Q000295:instrumentation / Q000379:methods; D053652:Nuclear Transfer Techniques / Q000295:instrumentation; D009865:Oocytes / Q000166:cytology*; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D014982:Xenopus laevis