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

合成生物学和系统生物学已经成为了解活细胞的关键研究领域。缺少的是一个实验系统，在这个实验系统中，来自全系统分析的知识可以被人工复制和研究。这个项目的目标是创造一个镜像合成生物学：即完全独立于自然的模拟生物系统，并从人造组成部分重新创造它。该项目将影响组学、系统生物学和合成生物学，并带来生命科学和化学的密切联系。人工系统将创造新的科学路线，并可能改变生物学的研究方式。同时，它将为生物技术和医学生产出具有独特性质和特征的分子。通过使用镜像成分的生物合成，更大镜像蛋白质的高质量生物合成可能成为现实。重要的是，一旦基本部件就位，机器本身就可以用来制造更多的部件。这最终将允许高质量地合成更大的镜像蛋白质。这些蛋白质应该是非免疫原性的，因此是新的蛋白质疗法和新的生物制造平台的潜在重要来源。技术摘要：在本项目中，研究人员将以L对映体核苷酸和D-氨基酸为原料，创建一个新型的镜像合成生物平台，用于合成功能镜像生物系统。几种基本的dna-dna、dna-蛋白质和蛋白质-蛋白质的相互作用以及几种酶的功能将从自然界复制到一个完全合成的以L-dna和D-蛋白质为基础的人工系统中。这将需要一些D型酶(dna连接酶、dna和rna聚合酶)和L dna序列以及形成基本对映体系统的辅助因子。此外，还将产生设计Ankyrin Repeat Proteins(DARPins)和转录因子或其DNA结合域的特定结合分子，并与其天然伙伴进行比较研究。除了人工生物学组件的建立，该项目还将立即追求实际用途，例如保护治疗性DARPins免受降解，因为它们不是天然蛋白酶的底物，例如允许口服应用，或创建SELEX选择程序从分子库中分离特定的L-DNA适配子。从长远来看，目标是建立一个自我复制的系统，最终包括D-蛋白质的生产。虽然这还很遥远，而且不会在这个项目中实现，但它将通过创建一个开发潜在非免疫原性镜像治疗蛋白的平台，在生物制造方面取得重大进展。

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.