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Genomic instability during reprogramming by nuclear transfer is DNA replication dependent

Nature Cell Biology volume 19, pages 282291 (2017) | Download Citation

Abstract

Somatic cells can be reprogrammed to a pluripotent state by nuclear transfer into oocytes, yet developmental arrest often occurs. While incomplete transcriptional reprogramming is known to cause developmental failure, reprogramming also involves concurrent changes in cell cycle progression and nuclear structure. Here we study cellular reprogramming events in human and mouse nuclear transfer embryos prior to embryonic genome activation. We show that genetic instability marked by frequent chromosome segregation errors and DNA damage arise prior to, and independent of, transcriptional activity. These errors occur following transition through DNA replication and are repaired by BRCA1. In the absence of mitotic nuclear remodelling, DNA replication is delayed and errors are exacerbated in subsequent mitosis. These results demonstrate that independent of gene expression, cell-type-specific features of cell cycle progression constitute a barrier sufficient to prevent the transition from one cell type to another during reprogramming.

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Acknowledgements

This research was supported by the New York Stem Cell Foundation (NYSCF) and a New York State Stem Cell Science (NYSTEM) IIRP Award no. C026184 and the John M. Driscoll, Jr, M. D. Children’s Fund. D.E. is a NYSCF-Robertson Investigator. We thank W. Kearns (Shady Grove Center for Preimplantation Genetic Diagnosis) for initial help with karyotyping. G.C. is supported by the Agency for Science, Technology and Research (A STAR) International Fellowship. We thank A. Ciccia and W.-W. Tee for critical comments on the manuscript.

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Affiliations

  1. Department of Pediatrics, Columbia University, New York 10032, USA

    • Gloryn Chia
    •  & Dieter Egli
  2. Department of Genetics and Genomic Sciences, Icahn School of Medicine at Mount Sinai, New York, New York 10029, USA

    • Judith Agudo
    •  & Brian D. Brown
  3. Reproductive Medicine Associates of New Jersey, New Jersey 07960, USA

    • Nathan Treff
  4. Center for Women’s Reproductive Care, College of Physicians and Surgeons, Columbia University, New York, New York 10019, USA

    • Mark V. Sauer
  5. Department of Obstetrics and Gynecology, College of Physicians and Surgeons, Columbia University, New York, New York 10032, USA

    • Mark V. Sauer
  6. Institute for Cancer Genetics, College of Physicians and Surgeons, Columbia University, New York 10032, USA

    • David Billing
    •  & Richard Baer

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Contributions

G.C. and D.E. designed and performed the experiments, analysed the data and wrote the manuscript with input from all authors. D.E. conceived and supervised the studies and performed NT. J.A. and B.D.B. provided T cells. N.T. performed karyotype analysis of blastomeres. D.B. and R.B. provided Brca1 mutant mice and helped with data interpretation and manuscript writing. M.V.S. was involved in all aspects of oocyte donation, including consent and oocyte retrieval.

Competing interests

The authors declare no competing financial interests.

Corresponding author

Correspondence to Dieter Egli.

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https://doi.org/10.1038/ncb3485

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