Viswanathan Ramya, Hoffman Elizabeth A, Shetty Savera J, Bekiranov Stefan, Auble David T
Department of Biochemistry and Molecular Genetics, University of Virginia Health System, Charlottesville, VA 22908, United States.
Department of Biochemistry and Molecular Genetics, University of Virginia Health System, Charlottesville, VA 22908, United States.
Methods. 2014 Dec;70(2-3):97-107. doi: 10.1016/j.ymeth.2014.10.029. Epub 2014 Nov 1.
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.
染色质中的转录因子结合位点通常通过染色质免疫沉淀测定法进行清点，这些结合模式可以提供有关细胞状态的精确而详细的信息。然而，一些关于转录因子功能的基本分子问题需要了解体内结合动力学以及位置信息。在这里，我们描述了交联动力学（CLK）测定法，其中甲醛交联的时间依赖性用于提取体内染色质结合的结合和解离速率。
