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Function of FEZF1 during early neural differentiation of human embryonic stem cells

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Abstract

The understanding of the mechanism underlying human neural development has been hampered due to lack of a cellular system and complicated ethical issues. Human embryonic stem cells (hESCs) provide an invaluable model for dissecting human development because of unlimited self-renewal and the capacity to differentiate into nearly all cell types in the human body. In this study, using a chemical defined neural induction protocol and molecular profiling, we identified Fez family zinc finger 1 (FEZF1) as a potential regulator of early human neural development. FEZF1 is rapidly up-regulated during neural differentiation in hESCs and expressed before PAX6, a well-established marker of early human neural induction. We generated FEZF1-knockout H1 hESC lines using CRISPR-CAS9 technology and found that depletion of FEZF1 abrogates neural differentiation of hESCs. Moreover, loss of FEZF1 impairs the pluripotency exit of hESCs during neural specification, which partially explains the neural induction defect caused by FEZF1 deletion. However, enforced expression of FEZF1 itself fails to drive neural differentiation in hESCs, suggesting that FEZF1 is necessary but not sufficient for neural differentiation from hESCs. Taken together, our findings identify one of the earliest regulators expressed upon neural induction and provide insight into early neural development in human.

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Acknowledgements

This work was supported by National Basic Research Program of China (2015CB964902 to Jiaxi Zhou and SQ2016ZY05002105 to Hongtao Wang), CAMS Initiative for Innovative Medicine (2016-I2M-1-018, 2016-I2M-3-002), National Natural Science Foundation of China (81530008, 31671541 to Jiaxi Zhou, 31500949 to Hongtao Wang), Tianjin Natural Science Foundation (16JCZDJC33100 to Jiaxi Zhou), and 3332015128 supported by PUMC Youth Fund and Fundamental Research Funds for the Central Universities to Dr. Hongtao Wang.

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Authors and Affiliations

  1. State Key Laboratory of Experimental Hematology, Institute of Hematology & Blood Diseases Hospital, Chinese Academy of Medical Sciences & Peking Union Medical College, Tianjin, 300020, China

    Xin Liu, Pei Su, Lisha Lu, Zicen Feng, Hongtao Wang & Jiaxi Zhou

  2. Center for Stem Cell Medicine, Chinese Academy of Medical Sciences & Department of Stem Cells and Regenerative Medicine, Peking Union Medical College, Tianjin, 300000, China

    Xin Liu, Pei Su, Lisha Lu, Zicen Feng, Hongtao Wang & Jiaxi Zhou

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  1. Xin Liu
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  2. Pei Su
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  3. Lisha Lu
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  4. Zicen Feng
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  5. Hongtao Wang
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  6. Jiaxi Zhou
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Correspondence to Hongtao Wang or Jiaxi Zhou.

Electronic supplementary material

Table S1

The primers used for real-time PCR

Table S2 The primers used for CRISPR sgRNA guide sequences and the genotyping

Table S3 The sources and dilutions of the antibodies

Supplementary material, approximately 27.5 KB.

Figure S1

Knockdown FEZF1 inhibited neural induction from hESCs.

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Liu, X., Su, P., Lu, L. et al. Function of FEZF1 during early neural differentiation of human embryonic stem cells. Sci. China Life Sci. 61, 35–45 (2018). https://doi.org/10.1007/s11427-017-9155-4

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