X chromosome inactivation in nuclear transfer ES cells.
Nuclear transfer ES (ntES) cells are established from cloned blastocysts generated by somatic cell nuclear transfer and are expected to be an important resource for regenerative medicine. However, cloned mammals, generated by similar methods, show various abnormalities, which suggest disordered gene regulation. Random X chromosome inactivation (XCI) has been observed to take place in cloned female mouse embryos, but XCI does not necessarily occur according to Xce strength, a genetic element that determines the likelihood of each X chromosome to be inactivated. This observation suggests incomplete reprogramming of epigenetic marks related to XCI. Here, we investigated XCI in ntES cell lines, which were established using differentiated embryoid bodies that originated from a female mouse ES cell line. We examined Xist RNA localization, histone modifications in the Xist locus, and XCI choice. We did not find substantial differences between the ntES lines and their parental ES line. This suggests that the Xist locus and the epigenetic marks involved in XCI are reprogrammed by nuclear transfer and subsequent ntES cell establishment. In contrast to skewed XCI in cloned mice, our observations indicate that normal XCI choice takes place in ntES cells, which supports the goal of safe therapeutic cloning for clinical use.
Shibata
S
S
Department of Stem Cell Biology, Graduate School of Medicine, Kanazawa University, Kanazawa, Japan. shinwa@med.kanazawa-u.ac.jp
Wakayama
T
T
Yokota
T
T
D000818:Animals; D001483:Base Sequence; D002460:Cell Line; D065150:Cellular Reprogramming / Q000235:genetics / Q000502:physiology; D047369:Chromatin Immunoprecipitation; D003582:Cytogenetics; D017931:DNA Primers / Q000235:genetics; D053595:Embryonic Stem Cells / Q000378:metabolism*; D005260:Female; D050172:Genes, X-Linked; D006657:Histones / Q000737:chemistry / Q000378:metabolism; D017404:In Situ Hybridization, Fluorescence; D051379:Mice; D053652:Nuclear Transfer Techniques; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014158:Transcription, Genetic; D049951:X Chromosome Inactivation / Q000235:genetics* / Q000502:physiology
D017931:DNA Primers; D006657:Histones