ERASynBio: Establishment of a Fully Synthetic, Mirror-Image Biological System

ERASynBio：建立全合成镜像生物系统

• 批准号: 1443228
• 负责人: Philip Dawson
• 金额: $ 55万
• 依托单位: The Scripps Research Institute
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-08-01 至 2017-07-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1443228&HistoricalAwards=false
• 关键词: ERASynBio; Establishment; Fully; Synthetic; Mirror

Synthetic Biology and Systems Biology have become key research areas in the quest for understanding living cells. What is missing is an experimental system in which the knowledge from system-wide analyses could be artificially reproduced and studied. The goal of this project is to create a mirror-image synthetic biology: that is, to mimic, entirely independent of nature, a biological system and to re-create it from artificial component parts. The project will influence -omics sciences, Systems Biology and Synthetic Biology and bring about a close contact of life sciences and chemistry. The artificial system will create new lines of science and may alter the way biology is done. At the same time, it will yield molecules of unique properties and characteristics for biotechnology and medicine. By using a biosynthesis from mirror components, a high-quality biosynthesis of larger mirror-image proteins might become a reality. Importantly, once the basic components are in place, the machinery itself can be used to create further components. This will eventually allow the high-quality synthesis of larger mirror image proteins. Such proteins should be non-immunogenic, and thus a potentially important source of new protein therapeutics and a new biomanufacturing platform. Technical Abstract: In this project, investigators will create a novel mirror-image synthetic biology platform from enantiomeric L-nucleotides and D-amino acids, which will be used to synthesize functional mirror-image biological system. Several basic DNA-DNA, DNA-protein and protein-protein interactions as well as the functions of few enzymes will be copied from nature into a totally synthetic artificial system based on L-DNA and D-proteins. This will require a few D-form enzymes (DNA-ligase, DNA- and RNA-polymerase) and L-DNA sequences plus co-factors that would form a basic enantiomeric system. In addition, particular binder molecules Designed Ankyrin Repeat Proteins (DARPins) and transcription factors, or DNA-binding domains thereof, will be generated and studied in comparison to their natural partners. Next to the establishment of components for artificial biology, there are immediate practical utilities that will be pursued within this project, such as the protection of therapeutic DARPins from degradation as they will not be substrates of natural proteases, permitting oral application for example, or the creation of a SELEX selection process for the isolation of specific L-DNA aptamers from a molecule library. In the long run, the aim is to set up a self-replicating system, eventually including D-protein production. While this is still way off and will not be achieved within this project, it would offer major advances in biomanufacturing, by creating a platform for the development of mirror image therapeutic proteins that are potentially non-immunogenic.

合成生物学和系统生物学已成为探索活细胞的关键研究领域。缺少的是一个实验系统，在这个系统中，可以人工复制和研究全系统分析的知识。该项目的目标是创建一个镜像合成生物学：也就是说，完全独立于自然，模仿生物系统，并从人工组成部分重新创建它。该项目将影响到组学、系统生物学和合成生物学，促进生命科学与化学的密切联系。人工系统将创造新的科学领域，并可能改变生物学的工作方式。与此同时，它将产生具有生物技术和医学独特性质和特征的分子。通过使用镜像组分的生物合成，较大镜像蛋白质的高质量生物合成可能成为现实。重要的是，一旦基本组件到位，机器本身就可以用来创建更多的组件。这最终将允许更大镜像蛋白质的高质量合成。 这些蛋白质应该是非免疫原性的，因此是新蛋白质治疗剂和新生物制造平台的潜在重要来源。 技术摘要：在本项目中，研究人员将从对映体L-核苷酸和D-氨基酸中创建一种新型镜像合成生物学平台，用于合成功能镜像生物系统。一些基本的DNA-DNA，DNA-蛋白质和蛋白质-蛋白质相互作用以及少数酶的功能将从自然界复制到基于L-DNA和D-蛋白质的完全合成的人工系统中。这将需要一些D型酶（DNA连接酶，DNA和RNA聚合酶）和L-DNA序列加上辅因子，形成基本的对映体系统。此外，将生成特定的结合分子设计的锚蛋白重复蛋白（DARPins）和转录因子或其DNA结合结构域，并与其天然伴侣进行比较研究。除了建立人工生物学组件之外，该项目还将追求直接的实用性，例如保护治疗性DARPin免于降解，因为它们不是天然蛋白酶的底物，例如允许口服应用，或者创建用于从分子文库中分离特定L-DNA适体的SELEX选择过程。从长远来看，目标是建立一个自我复制系统，最终包括D蛋白的生产。虽然这仍然很遥远，并且不会在该项目中实现，但它将通过创建一个开发潜在非免疫原性镜像治疗蛋白质的平台，为生物制造提供重大进展。