Analysis of Chromatin–Nuclear Receptor Interactions by Laser-Chromatin Immunoprecipitation

Rosaria Benedetti, Mariarosaria Conte, Vincenzo Carafa, Bartolomeo Della Ventura, Carlo Altucci, Raffaele Velotta, Hendrik G. Stunnenberg, Lucia Altucci, Angela Nebbioso

Research output: Contribution to journalArticlepeer-review

2 Citations (Scopus)

Abstract

Better defining the dynamics of biomolecular interactions is an important step in understanding molecular biology and cellular processes. DNA–protein interactions, and specifically hormone-triggered DNA– nuclear receptor interactions, are key events which are still poorly understood. To date, the most commonly used approach in studying chromatin interactions is the immunoprecipitation of chemically cross-linked chromatin (ChIP) coupled with single gene or global genomic analyses. Currently, establishing a stable interplay between nucleic acids and proteins (DNA–protein cross-link) is mainly obtained through conventional, diffusion-triggered, chemical methods using formaldehyde. Here we describe an alternative method, called Laser-ChIP (LChIP), for the specific analysis of interactions between chromatin and nuclear receptors driven by a UV laser energy source. Photo-induced cross-linking in LChIP is achieved very rapidly, allowing the study of transient interactions, depending on laser source parameters.

Original languageEnglish
Pages (from-to)25-34
Number of pages10
JournalMethods in molecular biology (Clifton, N.J.)
Volume1204
DOIs
Publication statusPublished - 2014
Externally publishedYes

Keywords

  • ChIP
  • Chromatin
  • Laser
  • Nuclear receptors
  • UV excitation

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