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、细胞周期和疾病（如癌症）中起重要作用。该提案有两个首要目标：（1）开发用于鉴定与特定DNA序列结合的蛋白质的方法，以及（2）开发用于鉴定针对复杂抗原源上发现的DNA结合蛋白质的免疫沉淀（IP）功能亲和试剂的更好方法。我们建议使用转录因子（TF）和它们的特异性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，从而使单个基因或多或少地活跃。虽然这是一个建议，以开发一种更好的手段，以获得功能性亲和试剂对蛋白质时，他们结合到DNA，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.