Ortt Kori, Sinha Satrajit
Department of Biochemistry, Center of Excellence in Bioinformatics and Life Sciences, State University of New York at Buffalo, Buffalo, NY, USA.
Methods Mol Biol. 2010;585:159-70. doi: 10.1007/978-1-60761-380-0_12.
Protein-DNA interactions, such as those that are necessary for transcription, are critical in regulating cellular function and behavior. The identification of DNA sequences that interact with transcriptional regulatory proteins is an important step necessary to better understand the molecular mechanisms regulating gene expression. Chromatin immunoprecipitation (ChIP) is one such procedure that provides a snapshot of which transcription factors are occupying specific DNA sequences. This method allows one not only to determine whether a particular genomic region is occupied by transcription factors but also to identify specific regulatory sequences that potentially control expression of their target genes. Recently, ChIP has been combined with both microarray analysis and a new generation of sequencing allowing a true genome-wide examination of transcription factor binding. Identifying the exact DNA sequence that a transcriptional regulatory protein binds, the precise timing of this association, and what other factors are involved in these interactions are important steps that will shed light on the transcriptional control mechanisms that dictate the biology of all cells, including keratinocytes.
蛋白质与DNA的相互作用，例如转录所必需的那些相互作用，在调节细胞功能和行为方面至关重要。识别与转录调节蛋白相互作用的DNA序列是更好地理解调节基因表达的分子机制所必需的重要一步。染色质免疫沉淀（ChIP）就是这样一种程序，它能提供哪些转录因子占据特定DNA序列的快照。这种方法不仅能让人确定特定基因组区域是否被转录因子占据，还能识别可能控制其靶基因表达的特定调节序列。最近，ChIP已与微阵列分析和新一代测序相结合，从而能够对转录因子结合进行真正的全基因组检查。确定转录调节蛋白所结合的确切DNA序列、这种结合的精确时间以及这些相互作用中涉及的其他因素，是重要的步骤，将有助于揭示决定所有细胞（包括角质形成细胞）生物学特性的转录控制机制。
