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High-throughput Identification of Gene Regulatory Sequences Using Next-generation Sequencing of Circular Chromosome Conformation Capture (4C-seq)
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Genetics
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JoVE Journal Genetics
High-throughput Identification of Gene Regulatory Sequences Using Next-generation Sequencing of Circular Chromosome Conformation Capture (4C-seq)

High-throughput Identification of Gene Regulatory Sequences Using Next-generation Sequencing of Circular Chromosome Conformation Capture (4C-seq)

DOI:
10.3791/58030-v

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09:06 min

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October 05, 2018

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, ,
1Division of Dermatology, Center for Pharmacogenomics, Center for the Study of Itch, Department of Medicine,Washington University School of Medicine

Chapters

  • 00:04Title
  • 00:45Formaldehyde Cross-linking and Cell Lysis
  • 01:59First Restriction Digestion and First Ligation
  • 05:07Cross-linking Reversion, Chromatin Isolation, and Further Analyses
  • 07:42Results: Representative Genomic Track of 4C-seq Read Coverage
  • 08:29Conclusion

Summary

Automatic Translation

Automatic Translation

The identification of physical interactions between genes and regulatory elements is challenging but has been facilitated by chromosome conformation capture methods. This modification to the 4C-seq protocol mitigates PCR bias by minimizing over-amplification of PCR templates and maximizes the mappability of reads by incorporating an addition restriction enzyme digest step.

Tags

Keywords: 4C-seqChromatin InteractionsGene Regulatory SequencesNext-generation SequencingTranscriptional RegulationPromoter-enhancer InteractionsCross-linkingRestriction DigestionCell Lysis

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