Cloning efficiency following ES cell nuclear transfer is influenced by the methylation state of the donor nucleus altered by mutation of DNA methyltransferase 3a and 3b |
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Authors: | Xiangpeng Dai Xiaoyang Zhao Hai Tang Jie Hao Jean-Paul Renard Qi Zhou Alice Jouneau Liu Wang |
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Institution: | 1.State Key Laboratory of Reproductive Biology, Institute of Zoology,Chinese Academy of Sciences,Beijing,China;2.Sino-French Laboratory LABIOCEM, Institute of Zoology,Chinese Academy of Sciences,Beijing,China;3.Graduate School of the Chinese Academy of Sciences,Beijing,China;4.INRA, UMR 1198 Biologie du Développement et Reproduction, Jouy en Josas,France ENVA,Maison-Alfort,France |
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Abstract: | The epigenetic state of donor cells plays a vital role in the nuclear reprogramming and chromatin remodeling of cloned embryos.
In this study we investigated the effect of DNA methylation state of donor cells on the development of mouse embryos reconstructed
with embryonic stem (ES) cell nuclei. Our results confirmed that deletion of the DNA methyltransferase 3a (Dnmt3a) and DNA
methyltransferase 3b (Dnmt3b) distinctly decreases the level of DNA methylation in ES cells. In contrast to wild type ES cells
(J1), Dnmt3a − / − 3b − / − (DKO) and Dnmt3b − / − (3bKO) donor cells significantly elevated the percentage of embryonic stem
cell nuclear transfer (ECNT) morula, blastocysts and postimplantation embryos (P < 0.05). However, the efficiency of establishment of NT-ES cell lines derived from DKO reconstructed blastocysts was not
improved, and the expression pattern of OCT4 and CDX2 in cloned blastocysts and postimplantation embryos was not altered either.
Our results suggest that the DNA methylation state of the donor nucleus is an important factor in regulation of the donor
nuclear reprogramming. |
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Keywords: | DNA methylation nuclear transfer DNA methyltransferase 3a DNA methyltransferase 3b |
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