Chromatin immunoprecipitation assays: Application of chip-on-chip for defining dynamic transcriptional mechanisms in bone cells

Margaretha Van Der Deen, Mohammad Q. Hassan, Jitesh Pratap, Nadiya M. Teplyuk, Daniel W. Young, Amjad Javed, Sayyed K. Zaidi, Jane B. Lian, Martin Montecino, Janet L. Stein, Gary S. Stein, Andre J. Van Wijnen

Resultado de la investigación: Contribución a los tipos de informe/libroCapítulorevisión exhaustiva

5 Citas (Scopus)

Resumen

Normal cell growth and differentiation of bone cells requires the sequential expression of cell type specific genes to permit lineage specification and development of cellular phenotypes. Transcriptional activation and repression of distinct sets of genes support the anabolic functions of osteoblasts and the catabolic properties of osteoclasts. Furthermore, metastasis of tumors to the bone environment is controlled by transcriptional mechanisms. Insights into the transcriptional regulation of genes in bone cells may provide a conceptual basis for improved therapeutic approaches to treat bone fractures, genetic osteopathologies, and/or cancer metastases to bone. Chromatin immunoprecipitation (ChIP) is a powerful technique to establish in vivo binding of transcription factors to the promoters of genes that are either activated or repressed in bone cells. Combining ChIP with genomic microarray analysis, colloquially referred to as "ChIP-on-chip," has become a valuable method for analysis of endogenous protein/DNA interactions. This technique permits assessment of chromosomal binding sites for transcription factors or the location of histone modifications at a genomic scale. This chapter discusses protocols for performing chromatin immunoprecipitation experiments, with a focus on ChIP-on-chip analysis. The information presented is based on the authors' experience with defining interactions of Runt-related (RUNX) transcription factors with bone-related genes within the context of the native nucleosomal organization of intact osteoblastic cells.

Idioma originalInglés
Título de la publicación alojadaOsteoporosis
Subtítulo de la publicación alojadaMethods and Protocols
EditorialHumana Press
Páginas165-176
Número de páginas12
ISBN (versión impresa)9781588298287
DOI
EstadoPublicada - 2008

Serie de la publicación

NombreMethods in Molecular Biology
Volumen455
ISSN (versión impresa)1064-3745

Áreas temáticas de ASJC Scopus

  • Biología molecular
  • Genética

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