Analysis of chromatin binding dynamics using the crosslinking kinetics (CLK) method

Methods. 2014 Dec;70(2-3):97-107. doi: 10.1016/j.ymeth.2014.10.029. Epub 2014 Nov 1.

Abstract

Transcription factor binding sites in chromatin are routinely inventoried by the chromatin immunoprecipitation assay, and these binding patterns can provide precise and detailed information about cell state. However, some fundamental molecular questions regarding transcription factor function require an understanding of in vivo binding dynamics as well as location information. Here we describe the crosslinking kinetics (CLK) assay, in which the time-dependence of formaldehyde crosslinking is used to extract on- and off-rates for chromatin binding in vivo.

Keywords: Chromatin immunoprecipitation; Dynamics; Formaldehyde; Kinetic; Method.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Binding Sites
  • Chromatin / chemistry*
  • Chromatin / metabolism
  • Chromatin Immunoprecipitation / methods
  • Formaldehyde / chemistry
  • Genetic Techniques*
  • Kinetics
  • Transcription Factors / chemistry*
  • Transcription Factors / metabolism
  • Yeasts / genetics

Substances

  • Chromatin
  • Transcription Factors
  • Formaldehyde