The GCE4All Center: Unleashing the Potential of Genetic Code Expansion for Biomedical Research

俄勒冈州拟建的GCE4所有生物医学技术开发和传播中心 大学(OSU)将优化、开发和广泛传播遗传密码扩展(GCE)技术- 细胞翻译工程，以表达含有非标准氨基酸(NCAA)的蛋白质。GCE 提供了前所未有的方法来探测和操纵大分子结构和功能，分析蛋白质 疾病中的故障，设计生物分析工具，创造新的精密生物疗法。GCE的可行性 已经很成熟了，但研究人员仍然很难访问和实施，因此仍然很少使用 -BTDD支持的理想目标。在其设想的≤15年寿命内，该中心的使命将是 优化和扩展现有的GCE技术，使非专业人员能够轻松使用，并广泛传播 通过广泛的教育、有效的培训以及提供可持续地获取优化技术 通过已建立的存储库-使强大的GCE方法成为标准的、广泛使用的工具 生物医学研究人员。创建拟议中心的优势在于我们在以下领域的经验和领先地位 开创性的前身OSU非天然蛋白质(UP)设施(2012-21)，其水平要小得多 开发和传播GCE方法并培训研究人员。GCE4所有的中心领导者都很好地- 准备通过其2个技术开发项目(TDP)、10个初步驱动来完成中心的使命 生物医学项目(DBP)和社区参与(CE)。协同式TDPS将优化和推广 GCE方法，用于1)使用GCE产生的蛋白质进行生物正交连接的应用，包括低- 在哺乳动物细胞中的背景标记和跟踪，以及2)产生含有生化的NCAA-蛋白质 探针和/或原生或模拟翻译后修饰(PTM)--无处不在，但鲜为人知 蛋白质功能的调节器。为了确保广泛的相关性，有针对性的技术进步将克服障碍 面对的是地理上不同的、由NIH资助的DBP，这些DBP将作为这项工作的严格试验床。要实现 中心优化将跨越4个阻碍研究人员的常见技术障碍 采用GCE。这4个GCE桥梁包括：用于1)纳入所选的NCAA的有效工具； 在2)大肠杆菌和3)哺乳动物细胞中高效地生产无杂质的GCE蛋白；以及4)产生稳定的 哺乳动物细胞系和细胞内可重复研究的可靠方案。行政长官核心将提供多样化的 培训活动，包括动手研讨会，已经被我们的UP设施证明是有效的。通过中心 网站上，CE将传播GCE方法、在线培训，并主办Wiki GCE知识库和GCE4所有 社区网络公告牌，在GCE用户社区中实现点对点支持。行政长官核心 还将组织两年一次的国际GCE会议，并确保所有优化的试剂公开 可从存储库中获取。GCE4所有中心将在1-5年内实现这些雄心勃勃的目标，以实现其 最终目标是将GCE从一种精品方法转变为分子生物学家工具箱的标准部分。

The proposed GCE4All Biomedical Technology Development and Dissemination Center at Oregon State University (OSU) will optimize, develop, and broadly disseminate Genetic Code Expansion (GCE) technology – the engineering of cellular translation to express proteins containing non-canonical amino acids (ncAAs). GCE provides unprecedented ways to probe and manipulate macromolecular structure and function, analyze protein malfunctions in disease, engineer bioanalytical tools, and create new precision biotherapeutics. GCE's feasibility is well-established, but it remains difficult for researchers to access and implement, and thus remains little-used – an ideal target for BTDD support. During its envisioned lifespan of ≤15 years, the Center's mission will be to optimize and extend existing GCE technologies to enable facile use by non-specialists, and to broadly disseminate them via widespread education, effective training, and by providing sustainable access to optimized technologies via established repositories – enabling powerful GCE approaches to become standard, widely-used tools of biomedical researchers. Advantageous for creating the proposed Center is our experience and leadership in the groundbreaking predecessor OSU Unnatural Protein (UP) Facility (2012-21), which at a much smaller level developed and disseminated GCE methods and trained researchers. GCE4All Center leaders are thus well- equipped to accomplish the Center mission via its 2 Technology Development Projects (TDPs), 10 initial Driving Biomedical Projects (DBPs), and Community Engagement (CE). The synergistic TDPs will optimize and extend GCE methods for 1) bioorthogonal ligation applications using GCE-produced proteins, including low- background labeling and tracking in mammalian cells, and 2) producing ncAA-proteins that contain biochemical probes and/or native or analog post-translational modifications (PTMs) – ubiquitous but little-understood regulators of protein functions. To ensure broad relevance, targeted technology advances will overcome barriers faced by geographically-diverse, NIH-funded DBPs that will serve as stringent testbeds for the work. To achieve its “GCE for All”goal, Center optimizations will bridge 4 common technological barriers that deter researchers from adopting GCE. These 4 GCE Bridges include: effective tools for 1) incorporating ncAAs of choice; for efficiently producing impurity-free GCE proteins in 2) E. coli and 3) mammalian cells; and 4) creation of stable mammalian cell lines and reliable protocols for reproducible studies in cells. The CE core will provide diverse training activities including hands-on workshops already proven effective by our UP Facility. Via the Center website, CE will disseminate GCE methods, online training, and host a Wiki GCE-knowledgebase and a GCE4All community networking bulletin board enabling peer-to-peer support in the GCE user community. The CE core will also organize biennial International GCE Conferences and ensure all optimized reagents are publicly available from repositories. The GCE4All Center will achieve these ambitious goals in Years 1-5, toward its ultimate goal of transforming GCE from a boutique method to a standard part of the molecular biologist's toolkit.