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

GCE4All 中心：释放遗传密码扩展在生物医学研究中的潜力

基本信息

批准号：
10335009
负责人：
RYAN A MEHL
金额：
$ 111.38万
依托单位：
OREGON STATE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-02-01 至 2027-01-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10335009
关键词：
Acquired Immunodeficiency Syndrome Adopted Amino Acids Automobile Driving Awareness Basic Science Beds Biochemical Biological Biological Process Biological Response Modifier Therapy Biomedical Research Biomedical Technology Biophysics Cancer Etiology Cardiovascular Diseases Cations Cell Line Cells Cellular Structures Chemicals Collaborations Communities Community Networks Development Diabetes Mellitus Diagnostic Diagnostic tests Disease Education Educational workshop Engineering Ensure Escherichia coli Evolution Foundations Funding Genetic Code Geography Goals Heart Diseases Inflammation International Knowledge Label Leadership Life Ligation Longevity Malignant Neoplasms Mammalian Cell Methods Mission Molecular Molecular Structure Nerve Degeneration Oregon Pain Parkinson Disease Pathogenesis Pharmaceutical Preparations Physiological Post-Translational Protein Processing Process Production Property Protein Dynamics Proteins Protocols documentation Reaction Reagent Regulation Reproducibility Research Research Personnel Site Specialist Speed System Technology Testing Tetanus Helper Peptide Therapeutic Training Training Activity Training Support Translations United States National Institutes of Health Universities Work analog base cellular engineering cohesion community engagement design experience functional group improved innovation interest knowledgebase peer peer support programs protein function protein structure function public repository repository symposium synergism technology development tool web site wiki

项目摘要

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.

拟议的俄勒冈州 GCE4All 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规范氨基酸 (ncAA) 的蛋白质。提供了前所未有的方法来探测和操纵大分子结构和功能，分析蛋白质疾病故障、设计生物分析工具并创造新的精准生物疗法的可行性。已经很成熟，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 ≤15 年的预期寿命内，该中心的使命是：优化和扩展现有的 GCE 技术，使非专业人士能够轻松使用，并广泛传播通过广泛的教育、有效的培训以及提供持续获得优化技术的机会来帮助他们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的优势在于我们在该领域的经验和领导力。开创性的前身 OSU 非天然蛋白质 (UP) 设施 (2012-21)，其规模要小得多因此，开发和传播 GCE 方法以及训练有素的研究人员是 GCE4All 中心的领导者。具备通过 2 个技术开发项目 (TDP)、10 个初始驱动来完成中心使命的能力生物医学项目 (DBP) 和社区参与 (CE) 协同 TDP 将优化和扩展。 GCE 方法用于 1) 使用 GCE 产生的蛋白质的生物正交连接应用，包括低哺乳动物细胞中的背景标记和追踪，以及 2) 产生含有生化物质的 ncAA 蛋白探针和/或天然或模拟翻译后修饰 (PTM) – 普遍存在但鲜为人知为了确保广泛的相关性，有针对性的技术进步将克服障碍。 NIH 资助的 DBP 面临着来自不同地区的挑战，这些 DBP 将作为这项工作的严格测试平台。其“GCE for All”目标，中心优化将弥合阻碍研究人员的 4 项常见技术障碍采用 GCE 的这 4 个 GCE 桥梁包括： 1) 纳入所选的 ncAA；在 2) 大肠杆菌和 3) 哺乳动物细胞中高效生产无杂质的 GCE 蛋白；以及 4) 建立稳定的 GCE 蛋白； CE 核心将提供哺乳动物细胞系和可靠的细胞可重复研究方案。培训活动（包括实践研讨会）已被我们的 UP 设施证明是有效的。网站，CE 将传播 GCE 方法、在线培训，并托管 Wiki GCE 知识库和 GCE4All 社区网络公告板，在 GCE 用户社区中提供点对点支持 CE 核心。还将组织两年一次的国际 GCE 会议，并确保所有优化试剂均公开 GCE4All 中心将在第 1-5 年实现这些雄心勃勃的目标，以实现其目标。最终目标是将 GCE 从精品方法转变为分子生物学家工具包的标准部分。