A challenge in developing proteolysis targeting chimeras (PROTACs) is the establishment of a universal platform applicable in multiple scenarios for precise degradation of proteins of interest (POIs). Inspired by the addressability, programmability, and rigidity of DNA frameworks, we develop covalent DNA framework-based PROTACs (DbTACs), which can be synthesized in high-throughput via facile bioorthogonal chemistry and self-assembly. DNA tetrahedra are employed as templates and the spatial position of each atom is defined. Thus, by precisely locating ligands of POI and E3 ligase on the templates, ligand spacings can be controllably manipulated from 8 Å to 57 Å. We show that DbTACs with the optimal linker length between ligands achieve higher degradation rates and enhanced binding affinity. Bispecific DbTACs (bis-DbTACs) with trivalent ligand assembly enable multi-target depletion while maintaining highly selective degradation of protein subtypes. When employing various types of warheads (small molecules, antibodies, and DNA motifs), DbTACs exhibit robust efficacy in degrading diverse targets, including protein kinases and transcription factors located in different cellular compartments. Overall, utilizing modular DNA frameworks to conjugate substrates offers a universal platform that not only provides insight into general degrader design principles but also presents a promising strategy for guiding drug discovery.

开发蛋白水解靶向嵌合体 (PROTAC) 的一个挑战是建立适用于多种场景的通用平台，以精确降解目标蛋白 (POI)。受 DNA 框架的可寻址性、可编程性和刚性的启发，我们开发了基于共价 DNA 框架的 PROTAC (DbTAC)，它可以通过简单的生物正交化学和自组装进行高通量合成。采用 DNA 四面体作为模板，定义每个原子的空间位置。因此，通过在模板上精确定位 POI 和 E3 连接酶的配体，配体间距可以在 8 Å 至 57 Å 范围内可控控制。我们表明，配体之间具有最佳连接长度的 DbTAC 可实现更高的降解率和增强的结合亲和力。具有三价配体组装的双特异性 DbTAC (bis-DbTAC) 能够实现多靶点去除，同时保持蛋白质亚型的高度选择性降解。当使用各种类型的弹头（小分子、抗体和 DNA 基序）时，DbTAC 在降解不同靶标方面表现出强大的功效，包括位于不同细胞区室的蛋白激酶和转录因子。总体而言，利用模块化 DNA 框架来缀合底物提供了一个通用平台，不仅提供了对一般降解剂设计原理的深入了解，而且还为指导药物发现提供了一种有前景的策略。