Functional-selection of affinity reagents against DNA-protein complexes using targeted chromatin sequences

描述（由申请人提供）：蛋白质与DNA的位点特异性结合在细胞发育、细胞信号传导、细胞周期和癌症等疾病中起着重要作用。本提案有两个主要目标：(1)开发一种方法来鉴定与特定DNA序列结合的蛋白质，(2)开发一种更好的方法来鉴定针对复杂抗原源上发现的DNA结合蛋白的免疫沉淀（IP）功能亲和试剂。我们建议使用转录因子（tf）及其特定DNA结合位点的相互作用作为模型来应用原理证明。tf是这种dna结合蛋白的一个主要例子。一旦结合到它们的结合位点，TF蛋白可以调节基因的转录，从而使单个基因的活性增加或减少。虽然这是一项建议，以开发一种更好的方法来获得针对蛋白质与DNA结合时的功能亲和试剂，但该方法本身可以应用于其他用途。

DESCRIPTION (provided by applicant): Site-specific binding of proteins to DNA plays an important role in cell development, cell signaling, the cell cycle, and diseases such as cancer. There are two overarching goals to this proposal: (1) to develop a means for the identification of proteins bound to specific DNA sequences, and (2) to develop a better method for the identification of immunoprecipitation (IP) functional affinity reagents against DNA-bound proteins found on a complex antigen source. We propose to apply the proof-of-principle using the interaction of transcription factors (TFs) and their specific DNA binding sites as a model. TFs are one major example of such DNA-binding proteins. Once bound to their binding site(s), TF proteins can regulate the transcription of genes, thereby making individual genes either more or less active. Although this is a proposal to develop a better means toward obtaining functional affinity reagents against proteins when they are bound to DNA, the method itself can be applied to other uses.