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    • 4. 发明授权
    • Assays for affinity profiling of nucleic acid binding proteins
    • 核酸结合蛋白亲和性分析的方法
    • US08932819B2
    • 2015-01-13
    • US13543711
    • 2012-07-06
    • Philip Dickinson
    • Philip Dickinson
    • G01N33/53C12Q1/68
    • G01N33/5308C12Q1/6806C12Q1/6811C12Q1/6827C12Q1/6837C12Q2522/101C12Q2533/101C12Q2565/525
    • Methods, compositions and kits are disclosed for assays to determine the binding affinity of DNA-binding proteins or RNA-binding proteins for their corresponding recognition site(s). In particular, assays are disclosed for measuring binding affinities when either the binding protein, or the recognition sequence of the recognition site, or cofactor proteins, contain one or more mutations. The disclosed assays can thus be utilized to measure the effect on transcription factor binding caused by mutations within the recognition site, or mutations within the binding domain of the protein, and to provide binding affinity information that can be correlated with altered gene regulation and expression. The disclosed assays can be personalized to a specific person or organism, with the measured binding affinities based upon an individual's specific binding proteins and recognition sites. Furthermore, embodiments are capable of measuring binding affinities between multiple binding proteins and multiple recognition sites through an entirely in vitro process.
    • 公开了用于测定DNA结合蛋白或RNA结合蛋白对于其相应识别位点的结合亲和力的方法,组合物和试剂盒。 特别地,公开了当结合蛋白或识别位点的识别序列或辅因子蛋白含有一个或多个突变时测量结合亲和力的测定。 因此,所公开的测定可用于测量由识别位点内的突变或蛋白质结合结构域内的突变引起的对转录因子结合的影响,并提供可与改变的基因调节和表达相关的结合亲和力信息。 所公开的测定可以针对特定人或生物进行个性化,其中所测量的结合亲和力基于个体的特异性结合蛋白和识别位点。 此外,实施方案能够通过完全体外方法测量多个结合蛋白和多个识别位点之间的结合亲和力。