title: Transient JMJD2B-mediated reduction of H3K9me3 levels improves reprogramming of embryonic stem cells into cloned embryos.
abstract: Correct reprogramming of epigenetic marks in the donor nuclei is crucial for successful cloning by nuclear transfer. Specific epigenetic modifications, such as repressive histone lysine methylation marks, are known to be very stable and difficult to reprogram. The discovery of histone lysine demethylases has opened up opportunities to study the effects of removing repressive histone lysine methylation marks in donor cells prior to nuclear transfer. In this study, we generated mouse embryonic stem (ES) cells for the inducible expression of JMJD2B (also known as KDM4B), a demethylase that primarily removes the histone-3 lysine-9 trimethylation (H3K9me3) mark. Induction of jmjd2b in the ES cells decreased total levels of H3K9me3 by 63%. When these cells were used for nuclear transfer, H3K9me3 levels were normalized within minutes following fusion with an enucleated oocyte. This transient reduction of H3K9me3 levels improved in vitro development into cloned embryos by 30%.
authors.affiliation: AgResearch Ruakura Research Centre, Hamilton, New Zealand.
authors.forename: Jisha
authors.initials: J
authors.lastname: Antony
authors.forename: Fleur
authors.initials: F
authors.lastname: Oback
authors.forename: Larry W
authors.initials: LW
authors.lastname: Chamley
authors.forename: Björn
authors.initials: B
authors.lastname: Oback
authors.forename: Götz
authors.initials: G
authors.lastname: Laible
mesh_terms: D000818:Animals; D000900:Anti-Bacterial Agents / Q000494:pharmacology; D002478:Cells, Cultured; D065150:Cellular Reprogramming; D019976:Cloning, Organism / Q000379:methods*; D004318:Doxycycline / Q000494:pharmacology; D053595:Embryonic Stem Cells / Q000166:cytology / Q000378:metabolism*; D005260:Female; D015870:Gene Expression / Q000187:drug effects; D049452:Green Fluorescent Proteins / Q000235:genetics; D006657:Histones / Q000378:metabolism*; D056484:Jumonji Domain-Containing Histone Demethylases / Q000235:genetics / Q000378:metabolism*; D008745:Methylation; D051379:Mice; D053652:Nuclear Transfer Techniques; D009865:Oocytes / Q000378:metabolism; D019076:Transgenes / Q000187:drug effects
chemical_list: D000900:Anti-Bacterial Agents; D006657:Histones; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins; D056484:Jumonji Domain-Containing Histone Demethylases; C511781:Kdm4b protein, mouse; D004318:Doxycycline