Cloned calves from chromatin remodeled in vitro.
We have developed a novel system for remodeling mammalian somatic nuclei in vitro prior to cloning by nuclear transplantation. The system involves permeabilization of the donor cell and chromatin condensation in a mitotic cell extract to promote removal of nuclear factors solubilized during chromosome condensation. The condensed chromosomes are transferred into enucleated oocytes prior to activation. Unlike nuclei of nuclear transplant embryos, nuclei of chromatin transplant embryos exhibit a pattern of markers closely resembling that of normal embryos. Healthy calves were produced by chromatin transfer. Compared with nuclear transfer, chromatin transfer shows a trend toward greater survival of cloned calves up to at least 1 mo after birth. This is the first successful demonstration of a method for directly manipulating the somatic donor chromatin prior to transplantation. This procedure should be useful for investigating mechanisms of nuclear reprogramming and for making improvements in the efficiency of mammalian cloning.
D000818:Animals; D015415:Biomarkers; D002417:Cattle; D002460:Cell Line; D002467:Cell Nucleus / Q000502:physiology*; D002843:Chromatin / Q000502:physiology*; D019976:Cloning, Organism / Q000379:methods*; D004247:DNA; D005314:Embryonic and Fetal Development; D005260:Female; D005347:Fibroblasts / Q000166:cytology; D066298:In Vitro Techniques; D034904:Lamin Type A / Q000378:metabolism; D009865:Oocytes / Q000502:physiology*; D011247:Pregnancy; D035181:TATA-Box Binding Protein / Q000378:metabolism
D015415:Biomarkers; D002843:Chromatin; D034904:Lamin Type A; D035181:TATA-Box Binding Protein; D004247:DNA