Anders Lars, Guenther Matthew G, Qi Jun, Fan Zi Peng, Marineau Jason J, Rahl Peter B, Lovén Jakob, Sigova Alla A, Smith William B, Lee Tong Ihn, Bradner James E, Young Richard A
1] Whitehead Institute for Biomedical Research, Cambridge, Massachusetts, USA. [2].
Department of Medical Oncology, Dana-Farber Cancer Institute, Massachusetts, USA.
Nat Biotechnol. 2014 Jan;32(1):92-6. doi: 10.1038/nbt.2776. Epub 2013 Dec 15.
A vast number of small-molecule ligands, including therapeutic drugs under development and in clinical use, elicit their effects by binding specific proteins associated with the genome. An ability to map the direct interactions of a chemical entity with chromatin genome-wide could provide important insights into chemical perturbation of cellular function. Here we describe a method that couples ligand-affinity capture and massively parallel DNA sequencing (Chem-seq) to identify the sites bound by small chemical molecules throughout the human genome. We show how Chem-seq can be combined with ChIP-seq to gain unique insights into the interaction of drugs with their target proteins throughout the genome of tumor cells. These methods will be broadly useful to enhance understanding of therapeutic action and to characterize the specificity of chemical entities that interact with DNA or genome-associated proteins.
大量小分子配体,包括正在研发和临床使用的治疗药物,通过与基因组相关的特定蛋白质结合来发挥其作用。在全基因组范围内绘制化学实体与染色质的直接相互作用的能力,可以为细胞功能的化学扰动提供重要见解。在此,我们描述了一种将配体亲和捕获与大规模平行DNA测序相结合的方法(化学测序法),以识别全人类基因组中被小分子化学物质结合的位点。我们展示了化学测序法如何与染色质免疫沉淀测序相结合,以深入了解药物在肿瘤细胞全基因组中与其靶蛋白的相互作用。这些方法将广泛有助于增进对治疗作用的理解,并表征与DNA或基因组相关蛋白质相互作用的化学实体的特异性。