| Chapter title |
Analysis of chromatin-nuclear receptor interactions by laser-chromatin immunoprecipitation.
|
|---|---|
| Chapter number | 3 |
| Book title |
Steroid Receptors
|
| Published in |
Methods in molecular biology, January 2014
|
| DOI | 10.1007/978-1-4939-1346-6_3 |
| Pubmed ID | |
| Book ISBNs |
978-1-4939-1345-9, 978-1-4939-1346-6
|
| Authors |
Rosaria Benedetti, Mariarosaria Conte, Vincenzo Carafa, Bartolomeo Della Ventura, Carlo Altucci, Raffaele Velotta, Hendrik G Stunnenberg, Lucia Altucci, Angela Nebbioso, Hendrik G. Stunnenberg |
| 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. |
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