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Abstract：
AS1411-NCL-MDM2-based proteolysis-targeting chimeras (ANM-PROTACs) are capable of inducing selective degradation of transcription factors (TFs) in tumor cells. Here, we present a protocol for constructing ANM-PROTACs. We describe steps for molecular design of the ANM-PROTACs, assembly and characterization of the ANM-PROTACs, and initial assessment of in vitro TF degradation potency. We then detail procedures for validation of selective degradation of TFs via proteomic analysis. This protocol has been successfully applied to degrade various TFs across multiple tumor cell lines. For complete details on the use and execution of this protocol, please refer to Fu et al.1.
摘要：
基于AS1411-NCL-MDM2的蛋白水解靶向嵌合体（ANM-PROTACs）能够诱导肿瘤细胞中转录因子（TF）的选择性降解。这里,我们提出了构建ANM-PROTACs的协议。我们描述了ANM-PROTACs的分子设计步骤，ANM-PROTACs的组装和表征，并初步评价了TF的体外降解效力。然后，我们详细说明了通过蛋白质组学分析验证TFs选择性降解的程序。该方案已成功应用于在多个肿瘤细胞系中降解各种TF。有关此协议的使用和执行的完整详细信息，PleaserefertoFuetal.1.
