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Targeted chromosome elimination from ES-somatic hybrid cells

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Abstract

To engineer a stem cell genome, we developed a technology for targeted elimination of chromosomes from mouse embryonic stem (ES)–somatic hybrid cells. Here we demonstrate the use of a universal chromosome elimination cassette (CEC) for elimination of a single embryonic stem cell (ESC)-derived chromosome 11 or 12, and also both copies of chromosome 6, which harbor pluripotency-associated genes including Nanog. We attribute hybrid-cell pluripotency to the expression of Nanog from the reprogrammed somatic-cell nuclei.

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Figure 1: ESCs carrying the CEC and cell fusion with somatic cells.
Figure 2: Chromosome painting of metaphase nuclei after elimination of chromosome 11 or 12.
Figure 3: Targeted elimination of a pair of ESC-derived chromosome 6s in a hybrid cell nucleus.

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Acknowledgements

We thank M. Evans for providing the HM1 ESC line, T. Shiroishi for providing the JF1 mice, L. Lefebvre for the pROSA26-pA and pROSA26-5′, A. McLaren and J. Ainscough for critical reading of the manuscript, and Y. Kurose for technical assistance.

Author information

Author notes

  1. Hiroyuki Matsumura and Masako Tada: These authors contributed equally to this work.

Authors and Affiliations

  1. Stem Cell Engineering, Institute for Frontier Medical Sciences, Kyoto University, 53 Kawahara-cho, Shogoin, Sakyo-ku, Kyoto, 606-8507, Japan

    Hiroyuki Matsumura, Masako Tada, Tomomi Otsuji & Takashi Tada

  2. ReproCELL Inc., 1-1-1 Uchisaiwai-cho, Chiyoda-ku, Tokyo, 100-0011, Japan

    Masako Tada & Tomomi Otsuji

  3. Development and Differentiation, Institute for Frontier Medical Sciences, Kyoto University, 53 Kawahara-cho, Shogoin, Sakyo-ku, Kyoto, 606-8507, Japan

    Kentaro Yasuchika & Norio Nakatsuji

  4. WellcomeTrust/Cancer Research UK Gurdon Institute of Cancer and Development Biology, University of Cambridge, Tennis Court Road, Cambridge, CB2 1QN, UK

    Azim Surani

  5. JST, CREST, 4-1-8 Hon-chou, Kawaguchi, Saitama, 332-0012, Japan

    Takashi Tada

Authors
  1. Hiroyuki Matsumura
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  2. Masako Tada
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  3. Tomomi Otsuji
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  4. Kentaro Yasuchika
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  5. Norio Nakatsuji
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  6. Azim Surani
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  7. Takashi Tada
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Corresponding author

Correspondence to Takashi Tada.

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The authors declare no competing financial interests.

Supplementary information

Supplementary Fig. 1

Chromosome elimination from CEC11 and CEC12 hybrid cells.

Supplementary Fig. 2

Karyotype of post-Cre CEC11 and CEC12 hybrid cells.

Supplementary Fig. 3

Targeted elimination of ESC-derived chromosome 6s.

Supplementary Fig. 4

Pluripotency of post-Cre CEC6tg/tg hybrid cells.

Supplementary Table 1

Chromosome elimination frequency.

Supplementary Methods

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Cite this article

Matsumura, H., Tada, M., Otsuji, T. et al. Targeted chromosome elimination from ES-somatic hybrid cells. Nat Methods 4, 23–25 (2007). https://doi.org/10.1038/nmeth973

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  • DOI: https://doi.org/10.1038/nmeth973

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