Abstract
Eukaryotic transcription is tightly regulated by transcriptional regulatory elements, even though these elements may be located far away from their target genes. It is now widely recognized that these regulatory elements can be brought in close proximity through the formation of chromatin loops, and that these loops are crucial for transcriptional regulation of their target genes. The chromosome conformation capture (3C) technique presents a snapshot of long-range interactions, by fixing physically interacting elements with formaldehyde, digestion of the DNA, and ligation to obtain a library of unique ligation products. Recently, several large-scale modifications to the 3C technique have been presented. Here, we describe chromosome conformation capture sequencing (4C-seq), a high-throughput version of the 3C technique that combines the 3C-on-chip (4C) protocol with next-generation Illumina sequencing. The method is presented for use in mammalian cell lines, but can be adapted to use in mammalian tissues and any other eukaryotic genome.
Author Contributions: Nele Gheldof and Alexandre Reymond wrote the paper. Marion Leleu, Daan Noordermeer, and Jacques Rougemont contributed the part about the data analysis.
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Acknowledgments
We are grateful for the help from Keith Harshman, Jérôme Thomas, Floriane Consales, and Emmanuel Beaudoing from the Lausanne Genomic Technologies Facility (GTF). This work was supported by the Marie Heim-Vögtlin fellowship to N.G.
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Gheldof, N., Leleu, M., Noordermeer, D., Rougemont, J., Reymond, A. (2012). Detecting Long-Range Chromatin Interactions Using the Chromosome Conformation Capture Sequencing (4C-seq) Method. In: Deplancke, B., Gheldof, N. (eds) Gene Regulatory Networks. Methods in Molecular Biology, vol 786. Humana Press. https://doi.org/10.1007/978-1-61779-292-2_13
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DOI: https://doi.org/10.1007/978-1-61779-292-2_13
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