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.
CITATION STYLE
Benedetti, R., Conte, M., Carafa, V., Ventura, B. D., Altucci, C., Velotta, R., … Nebbioso, A. (2014). Analysis of Chromatin–Nuclear Receptor Interactions by Laser-Chromatin Immunoprecipitation. Methods in Molecular Biology, 1204, 25–34. https://doi.org/10.1007/978-1-4939-1346-6_3
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