SAP-ERASER - Targeted Protein Degradation using SAP effectors

SAP-Eraser是一个雄心勃勃的高风险/高收益计划，旨在提供一种全新的靶向蛋白质降解(TPD)技术。SAP效应器是分泌的细菌蛋白，通过不同的机制介导宿主转录因子的降解。SAP05效应器直接结合26S蛋白酶体成分，导致不依赖E3连接酶和泛素化的蛋白质底物降解。SAP05的作用模式是独特的，因为其他蛋白降解靶向系统，如PROTAC，依赖底物泛素化和E3连接酶进行蛋白酶体依赖的降解。SAP05作为一种分子胶，夹在底物和26S蛋白酶体受体RPN10之间，导致底物的有效降解。因此，SAP05提供了一种独特的蛋白质敲除技术的机会，该技术在生物技术和生物医学中有许多应用，可以补充现有的基因敲除工具，如CRISPR-Cas和RNA干扰。在这里，我将结合生物物理、生化和遗传方法来确定SAP05效应器如何选择性地招募蛋白质进行降解，并开发具有新的底物结合特异性的合成SAP05变体。我围绕三个相互关联和互补的工作包(WPS)设计了这个项目，这三个工作包是生物物理、底物结合特异性和蛋白质降解技术。所获得的知识将作为开发SAP-橡皮擦技术的框架，将其作为一种有效的定向蛋白质降解工具，用于各种生物体的研究、治疗和生物技术应用，包括人类、动物、酵母和植物。

SAP-ERASER is an ambitious high-risk/high-gain program to deliver a radically novel Targeted Protein Degradation (TPD) technology. SAP effectors are secreted bacterial proteins that mediate the degradation of host transcription factors using different mechanisms. SAP05 effectors directly bind a 26S proteasome component leading to the degradation of protein substrates independently of E3 ligases and ubiquitination. SAP05 mode of action is unique because other proteolysis-targeting systems, such as PROTAC, depend on substrate ubiquitination and E3 ligases for proteasome-dependent degradation. SAP05 acts as a molecular glue that sandwiches a substrate and the 26S proteasome receptor RPN10 leading to efficient degradation of the substrate. Thus SAP05 offers the opportunity for a unique protein knock-down technology with many applications in biotechnology and biomedicine that complement existing gene knock-down tools, such as CRISPR-Cas and RNA interference. Here, I will combine biophysical, biochemical and genetic approaches to determine how SAP05 effectors selectively recruit proteins for degradation and to develop synthetic SAP05 variants with novel substrate binding specificities. I designed the project around three interconnected and complementary Work Packages (WPs), which are biophysics, substrate binding specificity and protein degradation technology. The gained knowledge will serve as a framework to develop the SAP-ERASER technology as an impactful targeted protein degradation tool for research, therapeutic and biotechnological applications in a variety of organisms, including humans, animals, yeast and plants.