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Identification of Potential Molecular Determinants of Murine Embryonic Stem Cell Differentiation by a Transposon-Based Approach

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Abstract

Embryonic stem cells (ESCs) are self-renewing pluripotent cells, capable of differentiating into all somatic cell types. The molecular control of self-renewal is relatively well-characterized, whereas how ESCs exit pluripotent state to differentiate is poorly understood. Here we identify two genes are required for differentiation and dozens of intergenic regions that potentially regulate ESC differentiation. We used PiggyBac (PB) transposon-based approach to randomly mutate the genome of ESCs, and generated hundreds of clones that resisted differentiation signals. Each clone was sequenced to determine genomic regions mutated by PB insertion. Intriguingly, many mutations were localized among intergenic regions and we identified two genes are required for differentiation. This study should facilitate further exploration of novel molecular determinants of embryonic stem cell differentiation.

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Acknowledgements

We thank Professor Pentao Liu (The Wellcome Trust Sanger Institute, United Kingdom) for kindly providing PiggyBac plasmid, Mrs. Ling Lin and the Science & Technology Department of West China Second University Hospital at Sichuan University for administrative assistance.

Funding

This work was supported by National Natural Science Foundation of China (81671115), and Science and Technology Department of Sichuan Province (2016JQ0029). The funders had no role in the study design, data collection and analysis, decision to publish, or preparation of the manuscript.

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Author notes

  1. Tong Qiu and Yin Fang have equally contributed to this work.

Authors and Affiliations

  1. Department of Pediatrics, West China Second University Hospital, Sichuan University, Chengdu, 610041, Sichuan, China

    Yan Wang, Tingjun Lei, Qian Dai, Tong Qiu & Yin Fang

  2. Center of Growth, Metabolism, and Aging, College of Life Sciences, Sichuan University, Chengdu, 610041, Sichuan, China

    Tingjun Lei

  3. Sichuan Cunde Therapeutics, Chengdu, 610093, China

    Ping Ding

  4. Key Laboratory of Birth Defects and Related Diseases of Women and Children, Ministry of Education, Sichuan University, Chengdu, 610041, Sichuan, China

    Yan Wang, Tingjun Lei, Qian Dai, Tong Qiu & Yin Fang

Authors
  1. Yan Wang
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  2. Tingjun Lei
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  3. Qian Dai
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  5. Tong Qiu
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Contributions

TQ and YF conceived the experiments. YW, TL, PD, and QD conducted experiments and participated in writing the manuscript. All authors reviewed the manuscript.

Corresponding authors

Correspondence to Tong Qiu or Yin Fang.

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The authors declare no competing financial interests.

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Wang, Y., Lei, T., Dai, Q. et al. Identification of Potential Molecular Determinants of Murine Embryonic Stem Cell Differentiation by a Transposon-Based Approach. Mol Biotechnol 60, 791–798 (2018). https://doi.org/10.1007/s12033-018-0110-7

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  • DOI: https://doi.org/10.1007/s12033-018-0110-7

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