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Single Cell Methods pp 251–267Cite as

Single-Cell 5fC Sequencing

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Part of the book series: Methods in Molecular Biology ((MIMB,volume 1979))

Abstract

Active DNA demethylation plays important roles in the epigenetic reprogramming of developmental processes. 5-formylcytosine (5fC) is produced during active demethylation of 5-methylcytosine (5mC). Here, we describe a technique called CLEVER-seq (Chemical-labeling-enabled C-to-T conversion sequencing), which detects the whole genome 5fC distribution at single-base and single-cell resolution. CLEVER-seq is suitable for the analysis of precious samples such as early embryos and laser microdissection captured samples.

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Acknowledgments

The authors thank Dr. Hongshan Guo, Bo Xia, Jinghui Song, and Hu Zeng for their help in developing the original protocol. Part of the analysis was performed on the Computing Platform of the Center for Life Sciences. This work was supported by the National Basic Research Program of China and the National Natural Science Foundation of China (91519325, MOST2016YFC0900300, 21522201, and 2014CB964900).

Competing financial interests: C.Z. and C.Y. are coinventors on filed patents (201710111600.9) for the labeling strategy and sequencing method reported herein.

Author information

Authors and Affiliations

  1. State Key Laboratory of Protein and Plant Gene Research, School of Life Sciences, Peking University, Beijing, People’s Republic of China

    Chenxu Zhu, Jinying Peng & Chengqi Yi

  2. Biodynamic Optical Imaging Center, Beijing Advanced Innovation Center for Genomics, School of Life Sciences, Peking University, Beijing, People’s Republic of China

    Yun Gao & Fuchou Tang

  3. Peking-Tsinghua Center for Life Sciences, Peking University, Beijing, People’s Republic of China

    Fuchou Tang & Chengqi Yi

  4. Ministry of Education Key Laboratory of Cell Proliferation and Differentiation, Peking University, Beijing, People’s Republic of China

    Fuchou Tang

  5. Department of Chemical Biology and Synthetic and Functional Biomolecules Center, College of Chemistry and Molecular Engineering, Peking University, Beijing, People’s Republic of China

    Chengqi Yi

Authors
  1. Chenxu Zhu
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  2. Yun Gao
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  3. Jinying Peng
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  4. Fuchou Tang
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Corresponding author

Correspondence to Chengqi Yi .

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Editors and Affiliations

  1. Department of Life Sciences and System Biology, University of Turin, Italian Institute for Genomic Medicine, IIGM Turin, Torino, Italy

    Valentina Proserpio

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Zhu, C., Gao, Y., Peng, J., Tang, F., Yi, C. (2019). Single-Cell 5fC Sequencing. In: Proserpio, V. (eds) Single Cell Methods. Methods in Molecular Biology, vol 1979. Humana, New York, NY. https://doi.org/10.1007/978-1-4939-9240-9_16

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  • DOI: https://doi.org/10.1007/978-1-4939-9240-9_16

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-4939-9239-3

  • Online ISBN: 978-1-4939-9240-9

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