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Detection of Copy Number Alterations Using Single Cell Sequencing
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Single-Cell 5fC Sequencing.

Chenxu Zhu1, Yun Gao2, Jinying Peng1

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

Methods in Molecular Biology (Clifton, N.J.)
|April 28, 2019
PubMed
Summary
This summary is machine-generated.

We developed CLEVER-seq, a new method to map the whole genome distribution of 5-formylcytosine (5fC) at single-cell resolution. This technique is crucial for understanding epigenetic reprogramming in precious samples like early embryos.

Keywords:
5-FormylcytosineBisulfite-free sequencingCLEVER-seqChemical labelingSingle cell

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Area of Science:

  • Epigenetics and Genomics
  • Molecular Biology
  • Developmental Biology

Background:

  • Active DNA demethylation is a key epigenetic process.
  • 5-formylcytosine (5fC) is an intermediate in the active demethylation of 5-methylcytosine (5mC).
  • Understanding 5fC distribution is vital for studying epigenetic reprogramming.

Purpose of the Study:

  • To introduce a novel technique for genome-wide 5fC detection.
  • To enable single-base and single-cell resolution analysis of 5fC.
  • To facilitate the study of epigenetic dynamics in limited or precious biological samples.

Main Methods:

  • Development of CLEVER-seq (Chemical-labeling-enabled C-to-T conversion sequencing).
  • Chemical labeling of 5fC followed by C-to-T conversion.
  • Whole-genome sequencing to determine 5fC distribution at single-base resolution.
  • Application to single-cell samples.

Main Results:

  • CLEVER-seq successfully detects genome-wide 5fC distribution.
  • The method achieves single-base and single-cell resolution.
  • Demonstrated suitability for analyzing precious samples, including early embryos.

Conclusions:

  • CLEVER-seq provides a powerful tool for high-resolution mapping of 5fC.
  • This technique advances the study of active DNA demethylation dynamics.
  • Enables new insights into epigenetic reprogramming during development and in rare cell populations.