Stem Cell Reports (Feb 2018)

Reduced Self-Diploidization and Improved Survival of Semi-cloned Mice Produced from Androgenetic Haploid Embryonic Stem Cells through Overexpression of Dnmt3b

  • Wenteng He,
  • Xiaobai Zhang,
  • Yalin Zhang,
  • Weisheng Zheng,
  • Zeyu Xiong,
  • Xinjie Hu,
  • Mingzhu Wang,
  • Linfeng Zhang,
  • Kun Zhao,
  • Zhibin Qiao,
  • Weiyi Lai,
  • Cong Lv,
  • Xiaochen Kou,
  • Yanhong Zhao,
  • Jiqing Yin,
  • Wenqiang Liu,
  • Yonghua Jiang,
  • Mo Chen,
  • Ruimin Xu,
  • Rongrong Le,
  • Chong Li,
  • Hong Wang,
  • Xiaoping Wan,
  • Hailin Wang,
  • Zhiming Han,
  • Cizhong Jiang,
  • Shaorong Gao,
  • Jiayu Chen

Journal volume & issue
Vol. 10, no. 2
pp. 477 – 493

Abstract

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Summary: Androgenetic haploid embryonic stem cells (AG-haESCs) hold great promise for exploring gene functions and generating gene-edited semi-cloned (SC) mice. However, the high incidence of self-diploidization and low efficiency of SC mouse production are major obstacles preventing widespread use of these cells. Moreover, although SC mice generation could be greatly improved by knocking out the differentially methylated regions of two imprinted genes, 50% of the SC mice did not survive into adulthood. Here, we found that the genome-wide DNA methylation level in AG-haESCs is extremely low. Subsequently, downregulation of both de novo methyltransferase Dnmt3b and other methylation-related genes was determined to be responsible for DNA hypomethylation. We further demonstrated that ectopic expression of Dnmt3b in AG-haESCs could effectively improve DNA methylation level, and the high incidence of self-diploidization could be markedly rescued. More importantly, the developmental potential of SC embryos was improved, and most SC mice could survive into adulthood. : Ectopic expression of Dnmt3b could rescue DNA methylation level in repetitive sequences of hypomethylated AG-haESCs, suppress high incidence of self-diploidization, and promote developmental potential of SC embryos, and most SC mice could survive into adulthood. Keywords: androgenetic haploid embryonic stem cells, self-diploidization, semi-cloned mice, DNA methylation, Dnmt3b