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Peer-reviewedCC BY 4.0STAR Protocols

Protocol to identify DNA methylation-sensitive enhancer sites

  • Natalie Taege1
  • Sonja C. Schriever2
  • Henriette Kirchner1
  1. 1Institute for Human Genetics, Division Epigenetics & Metabolism, University of Luebeck, Ratzeburger Allee 160, 23562 Luebeck, Germany
  2. 2Research Unit NeuroBiology of Diabetes, Institute for Diabetes and Obesity, Helmholtz Centre Munich, Ingolstädter Landstrasse 1, 85764 Neuherberg, Germany
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Abstract

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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Cite

BibTeXRIS
BibTeX
@article{taege2026protocol,
  title = {{Protocol to identify DNA methylation-sensitive enhancer sites}},
  author = {Natalie Taege and Sonja C. Schriever and Henriette Kirchner},
  journal = {STAR Protocols},
  year = {2026},
  month = oct,
  doi = {10.1016/j.xpro.2026.104922},
  url = {https://doi.org/10.1016/j.xpro.2026.104922},
  note = {CC BY 4.0},
}
RIS
TY  - JOUR
TI  - Protocol to identify DNA methylation-sensitive enhancer sites
AU  - Natalie Taege
AU  - Sonja C. Schriever
AU  - Henriette Kirchner
PY  - 2026
DA  - 2026/10/29
JO  - STAR Protocols
DO  - 10.1016/j.xpro.2026.104922
UR  - https://doi.org/10.1016/j.xpro.2026.104922
AB  - 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
SN  - 2666-1667
ER  -