Assessing sites of NF-κB DNA binding using chromatin immunoprecipitation.

Colleran, A., Collins, P. E. and Carmody, R. J. (2015) Assessing sites of NF-κB DNA binding using chromatin immunoprecipitation. In: May, M.J. (ed.) NF-kappaB Methods and Protocols. Series: Methods in molecular biology (1280). Humana Press, pp. 47-59. ISBN 9781493924226 (doi: 10.1007/978-1-4939-2422-6_4)

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Abstract

The NF-κB transcription factor is in fact a family of related proteins which dimerize to form at least 12 distinct complexes which regulate the expression of hundred of genes of importance to a range of physiological and pathological processes. The binding of NF-κB to the regulatory regions and promoters of target genes is influenced by a number of factors including the sequence of DNA-binding sites, the posttranslational modification of NF-κB, and the interaction of cofactors and co-regulators of transcription. In addition, the binding of NF-κB to promoters is highly dynamic and the recruitment of specific subunits to specific binding sites may occur with distinct kinetics. Moreover, genome-wide analysis of NF-κB chromatin binding indicates that the majority of DNA-binding events are not associated with changes in transcriptional activity. Thus, the analysis of NF-κB recruitment and activity at specific binding sites is of critical importance in understanding the regulation of transcription. In this chapter we describe a chromatin immunoprecipitation assay to investigate the in situ binding of NF-κB to specific sites in the genome.

Item Type:Book Sections
Status:Published
Glasgow Author(s) Enlighten ID:Carmody, Dr Ruaidhri and Collins, Ms Patricia
Authors: Colleran, A., Collins, P. E., and Carmody, R. J.
Subjects:Q Science > QH Natural history > QH345 Biochemistry
College/School:College of Medical Veterinary and Life Sciences > School of Infection & Immunity
Journal Name:Methods in molecular biology (Clifton, N.J.)
Publisher:Humana Press
ISSN:1940-6029
ISBN:9781493924226
Published Online:01 February 2015

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