期刊
NATURE PROTOCOLS
卷 13, 期 3, 页码 551-564出版社
NATURE PORTFOLIO
DOI: 10.1038/nprot.2017.150
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资金
- University of Cambridge
- Cancer Research UK [C14303/A17197]
- Wellcome Trust [099232/z/12/z]
- European Union [747297-QAPs-H2020-MSCA-IF-2016]
- Cancer Research UK [19836] Funding Source: researchfish
G-rich DNANA sequences can form four-stranded G-quadruplex (G4) secondary structures and are linked to fundamental biological processes such as transcription, replication and telomere maintenance. G4s are also implicated in promoting genome instability, cancer and other diseases. Here, we describe a detailed G4 ChIP-seq method that robustly enables the determination of G4 structure formation genome-wide in chromatin. This protocol adapts traditional ChIP-seq for the detection of DNANA secondary structures through the use of a G4-structure-specific single-chain antibody with refinements in chromatin immunoprecipitation followed by high-throughput sequencing. This technology does not require expression of the G4 antibody in situ, enabling broad applicability to theoretically all chromatin sources. Beginning with chromatin isolation and antibody preparation, the entire protocol can be completed in < 1 week, including basic computational analysis.
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