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Protocol to identify DNA methylation-sensitive enhancer sites.
STAR Protoc. 7:104922 (2026)
Here, we present a protocol to identify DNA methylation-sensitive enhancer sites using the leptin-regulating enhancer RS1 in adipose tissue. We describe steps for validating RS1's endogenous function with a dCas9-KRAB repressor system and quantifying RS1 CpG methylation by pyrosequencing. We then detail procedures for assessing methylation sensitivity by luciferase reporter assay. This protocol is adaptable to other enhancers and tissues. For complete details on the use and execution of this protocol, please refer to Taege et al.1.
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Publication type
Article: Journal article
Document type
Review
Keywords
Dna Methylation ; Enhancers ; Enhancer Identification ; Epigenetics ; Chromatin Regulation
e-ISSN
2666-1667
Journal
STAR Protocols
Quellenangaben
Volume: 7,
Issue: 4,
Article Number: 104922
Publisher
Elsevier
Reviewing status
Peer reviewed
Institute(s)
Institute of Diabetes and Obesity (IDO)