TY - JOUR
T1 - Analysis of Chromatin–Nuclear Receptor Interactions by Laser-Chromatin Immunoprecipitation
AU - Benedetti, Rosaria
AU - Conte, Mariarosaria
AU - Carafa, Vincenzo
AU - Ventura, Bartolomeo Della
AU - Altucci, Carlo
AU - Velotta, Raffaele
AU - Stunnenberg, Hendrik G.
AU - Altucci, Lucia
AU - Nebbioso, Angela
N1 - Publisher Copyright:
© Springer Science+Business Media New York 2014.
PY - 2014
Y1 - 2014
N2 - 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.
AB - 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.
KW - ChIP
KW - Chromatin
KW - Laser
KW - Nuclear receptors
KW - UV excitation
UR - http://www.scopus.com/inward/record.url?scp=84921826040&partnerID=8YFLogxK
U2 - 10.1007/978-1-4939-1346-6_3
DO - 10.1007/978-1-4939-1346-6_3
M3 - Article
C2 - 25182758
AN - SCOPUS:84921826040
SN - 1940-6029
VL - 1204
SP - 25
EP - 34
JO - Methods in molecular biology (Clifton, N.J.)
JF - Methods in molecular biology (Clifton, N.J.)
ER -