FiTAc-seq: fixed-tissue ChIP-seq for H3K27ac profiling and super-enhancer analysis of FFPE tissues.
Nat Protoc
; 15(8): 2503-2518, 2020 08.
Article
em En
| MEDLINE
| ID: mdl-32591768
ABSTRACT
Fixed-tissue ChIP-seq for H3K27 acetylation (H3K27ac) profiling (FiTAc-seq) is an epigenetic method for profiling active enhancers and promoters in formalin-fixed, paraffin-embedded (FFPE) tissues. We previously developed a modified ChIP-seq protocol (FiT-seq) for chromatin profiling in FFPE. FiT-seq produces high-quality chromatin profiles particularly for methylated histone marks but is not optimized for H3K27ac profiling. FiTAc-seq is a modified protocol that replaces the proteinase K digestion applied in FiT-seq with extended heating at 65 °C in a higher concentration of detergent and a minimized sonication step, to produce robust genome-wide H3K27ac maps from clinical samples. FiTAc-seq generates high-quality enhancer landscapes and super-enhancer (SE) annotation in numerous archived FFPE samples from distinct tumor types. This approach will be of great interest for both basic and clinical researchers. The entire protocol from FFPE blocks to sequence-ready library can be accomplished within 4 d.
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Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Histonas
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Fixação de Tecidos
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Inclusão em Parafina
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Sequenciamento de Cromatina por Imunoprecipitação
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Lisina
Tipo de estudo:
Guideline
Limite:
Animals
Idioma:
En
Ano de publicação:
2020
Tipo de documento:
Article