ERA SynBio: Design and Synthesis of a Bio-orthogonal Genetic System

合成生物学作为一门能够回答基本生物学问题并为许多实际应用做出贡献的科学学科，前景广阔。这一领域的一个关键障碍是对工程有机体逃逸到环境中的担忧。为了解决这一问题和其他担忧，这个由美国、英国、法国和西班牙的研究人员组成的研究联盟将设计一种由非天然核酸组成的独立复制元件(称为Episome，它不整合到宿主细胞的DNA中)。这些episome将拥有基于TNA(基于苏糖的核酸)的遗传物质，以及一个复制和表达Episome蛋白质的系统。自主的以TNA为基础的Episome不能在自然界(实验室外)繁殖，因为TNA前体不是自然产生的。每个研究人员联盟都将开发特定的成分，将其整合到TNA Episome中，并将其转移到细菌宿主中。该研究项目虽然风险很高，但预计将在提供使能技术以推进合成生物学启发的生物经济方面产生重大影响。它还将对遗传物质的进化产生洞察力。该项目的培训潜力非同寻常，因为学生和初级研究人员将获得有价值的跨学科和国际培训。该项目的目标是通过开发基于噬菌体Phi29的全TNA Episome来重建分子生物学的中心教条，该Episome可以在体外复制，并最终使用为此设计的正交细胞机械在活细胞内复制。这个工程系统将蛋白质编码信息存储在TNA聚合物中，这种聚合物将独立于细胞基因组进行复制。TNA编码信息将由一种进化的RNA聚合酶转录，该聚合酶可以识别TNA模板并产生天然信使RNA，用于翻译成功能蛋白质。该项目由美国国家科学基金会和欧洲资助机构之间的跨国资助机制资助，这些机构是欧洲委员会批准的合成生物学研究区域网络的一部分。该项目的美国部分由系统和合成生物学(生物科学局)和生物技术和生化生物工程(工程局)方案共同资助。

Synthetic biology holds great promise as a scientific discipline that can answer fundamental biological questions as well as contribute to many practical applications. A key obstacle for this field is the concern about escape of an engineered organism into the environment. To address this and other concerns, this research consortium that engages researchers in the US, UK, France and Spain will engineer an independently replicating element (called an episome, which is not integrated into the DNA of the host cell) that consists of non-natural nucleic acids. These episomes will have genetic material based on TNA (threose-based nucleic acid) along with a system that replicates and expresses proteins from the episome. An autonomous TNA-based episome could not be propagated in nature (outside the laboratory) because TNA precursors do not occur naturally. The consortium of investigators each will develop specific components to be incorporated into the TNA episome, which will be transferred into a bacterial host. The research project, while high risk, is anticipated to generate high impacts in terms of providing enabling technologies to advance a synthetic biology-inspired bioeconomy. It will also generate insights into the evolution of genetic material. The training potential of this project is exceptional in that the students and junior researchers will gain valuable transdisciplinary and international training.The goal of this project is to re-structure the central dogma of molecular biology by developing an all-TNA episome based on bacteriophage Phi29 that can replicate in vitro and eventually inside living cells using an orthogonal cellular machinery engineered for this purpose. The engineered system will store protein-coding information in TNA polymers that will replicate independently of the cellular genome. The TNA coding information will be transcribed by an evolved RNA polymerase that can recognize the TNA template and produce natural messenger RNAs for translation into functional proteins.This project is funded through a transnational funding mechanism between the United States National Science Foundation and European Funding Agencies that are part of the European Commission endorsed Research Area Network in Synthetic Biology. The United States component of this project is co-funded by programs in Systems and Synthetic Biology (Directorate for Biological Sciences) and Biotechnology and Biochemical Bioengineering (Directorate for Engineering).