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Cell Stem Cell. 2018 Sep 6;23(3):343-354.e5. doi: 10.1016/j.stem.2018.06.008. Epub 2018 Jul 19.

Loss of H3K27me3 Imprinting in Somatic Cell Nuclear Transfer Embryos Disrupts Post-Implantation Development.

Author information

1
Howard Hughes Medical Institute; Program in Cellular and Molecular Medicine, Boston Children's Hospital, Boston, MA 02115, USA; Department of Genetics, Harvard Medical School, Boston, MA 02115, USA; RIKEN Bioresource Research Center, Tsukuba, Ibaraki 305-0074, Japan; Cooperative Division of Veterinary Sciences, Tokyo University of Agriculture and Technology, Fuchu, Tokyo 183-8509, Japan.
2
Life Sciences Institute and Stem Cell Institute, Zhejiang University, Hangzhou 310058, China.
3
Howard Hughes Medical Institute; Program in Cellular and Molecular Medicine, Boston Children's Hospital, Boston, MA 02115, USA; Department of Genetics, Harvard Medical School, Boston, MA 02115, USA.
4
RIKEN Bioresource Research Center, Tsukuba, Ibaraki 305-0074, Japan.
5
Howard Hughes Medical Institute; Department of Molecular Biology, Massachusetts General Hospital, Boston, MA 02115, USA; Department of Genetics, Harvard Medical School, Boston, MA 02115, USA.
6
RIKEN Bioresource Research Center, Tsukuba, Ibaraki 305-0074, Japan; RIKEN Cluster for Pioneering Research, Wako, Saitama 351-0198, Japan.
7
Howard Hughes Medical Institute; Program in Cellular and Molecular Medicine, Boston Children's Hospital, Boston, MA 02115, USA; Department of Genetics, Harvard Medical School, Boston, MA 02115, USA; Life Sciences Institute and Stem Cell Institute, Zhejiang University, Hangzhou 310058, China. Electronic address: li_shen@zju.edu.cn.
8
Howard Hughes Medical Institute; Program in Cellular and Molecular Medicine, Boston Children's Hospital, Boston, MA 02115, USA; Harvard Stem Cell Institute, Boston, MA 02115, USA; Department of Genetics, Harvard Medical School, Boston, MA 02115, USA. Electronic address: yzhang@genetics.med.harvard.edu.

Abstract

Animal cloning can be achieved through somatic cell nuclear transfer (SCNT), although the live birth rate is relatively low. Recent studies have identified H3K9me3 in donor cells and abnormal Xist activation as epigenetic barriers that impede SCNT. Here we overcome these barriers using a combination of Xist knockout donor cells and overexpression of Kdm4 to achieve more than 20% efficiency of mouse SCNT. However, post-implantation defects and abnormal placentas were still observed, indicating that additional epigenetic barriers impede SCNT cloning. Comparative DNA methylome analysis of IVF and SCNT blastocysts identified abnormally methylated regions in SCNT embryos despite successful global reprogramming of the methylome. Strikingly, allelic transcriptomic and ChIP-seq analyses of pre-implantation SCNT embryos revealed complete loss of H3K27me3 imprinting, which may account for the postnatal developmental defects observed in SCNT embryos. Together, these results provide an efficient method for mouse cloning while paving the way for further improving SCNT efficiency.

KEYWORDS:

DNA methylation; H3K27me3-dependent imprinting; animal cloning; epigenetic reprogramming; genomic imprinting; mouse embryo; somatic cell nuclear transfer

PMID:
30033120
DOI:
10.1016/j.stem.2018.06.008

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