Cell-free synthesis of iron-sulfur clusters for artificial cells

用于人造细胞的铁硫簇的无细胞合成

• 批准号: RGPIN-2020-04375
• 负责人: Mansy, Sheref
• 金额: $ 6.85万
• 依托单位: University of Alberta
• 依托单位国家: 加拿大
• 项目类别: Discovery Grants Program - Individual
• 财政年份: 2021
• 资助国家: 加拿大
• 起止时间: 2021-01-01 至 2022-12-31
• 项目状态: 已结题
• 来源: https://www.nserc-crsng.gc.ca/ase-oro/Details-Detailles_eng.asp?id=739930
• 关键词: Cell; free; synthesis; iron; sulfur

Artificial cells are mimics of living cells built in the laboratory with DNA, protein, and lipids. Although building such systems in the laboratory is more challenging than engineering living cells, there are numerous benefits. First, there is arguably no better way to understand how biology works than to try to build biological systems from component parts. Further, we can intentionally put in the parts of biology that are advantageous when building a cellular mimic from scratch while simultaneously leaving out that problematic parts of biology. For example, non-living cellular mimics, i.e. artificial cells, can be built to not replicate, which completely removes the fear of losing control of the system. There can be no long-term ecological consequences, because once the artificial cells fall apart, the environment returns to its prior state. We have built artificial cells that can chemically communicate with and control natural living cells. Such technologies have numerous potential applications, from cleaning our environment of pollutants to treating debilitating diseases and infections. However, the state-of-the-art in artificial cells are not long-lived. That is, the artificial cells are active for a couple of hours then degrade. Here, we intend to give the artificial cells metabolism so that the artificial cells can function long enough to be useful. To do so, we need to learn how to build the components of metabolism outside of a living cell. We will start by tackling the synthesis of one of the more complex parts of metabolism, which is the synthesis of iron-sulfur proteins. Iron-sulfur clusters are ancient, inorganic arrangements of atoms that mediate much of the chemistry that keeps cells alive. By exploiting our expertise in the synthesis of iron-sulfur clusters and in in vitro genetics, we will develop a genetically encoded pathway within artificial cells that synthesizes the iron-sulfur proteins of metabolism. These proteins will then be combined with other genetically encoded factors to begin giving artificial cells a rudimentary form of metabolism. Additionally, we will exploit the developed pathways to generate new types of catalysts capable of producing biofuels. Both iron-sulfur protein and iron-sulfur peptide catalysts will be pursued for potential applications in industry. We are one of the leaders in the synthesis of artificial cells. This project will help us consolidate our position, potentially giving rise to the generation of new technologies and companies. Additionally, this project will enrich our understanding of the chemical foundations of biology.

人工细胞是在实验室中用DNA、蛋白质和脂质构建的活细胞的模拟物。虽然在实验室中构建这样的系统比工程活细胞更具挑战性，但有许多好处。首先，要理解生物学是如何工作的，没有比尝试从组成部分构建生物系统更好的方法了。此外，我们可以有意识地加入生物学中对从头开始构建细胞模拟物有利的部分，同时忽略生物学中有问题的部分。例如，非活体细胞模拟物，即人工细胞，可以被构建为不复制，这完全消除了对失去系统控制的恐惧。不会产生长期的生态后果，因为一旦人造细胞崩溃，环境就会恢复到之前的状态。我们已经建立了人造细胞，可以与天然活细胞进行化学交流并控制它们。这些技术有许多潜在的应用，从清洁我们环境中的污染物到治疗使人衰弱的疾病和感染。然而，最先进的人造细胞都不是长寿的。也就是说，人工细胞在几个小时内是活跃的，然后降解。在这里，我们打算给人工细胞新陈代谢，使人工细胞可以发挥作用足够长的时间是有用的。要做到这一点，我们需要学习如何在活细胞外构建新陈代谢的组成部分。我们将从处理代谢中更复杂部分之一的合成开始，即铁硫蛋白的合成。铁硫簇是古老的无机原子排列，介导了许多维持细胞存活的化学物质。通过利用我们在铁硫簇合成和体外遗传学方面的专业知识，我们将在人工细胞内开发一种遗传编码途径，合成代谢的铁硫蛋白。然后这些蛋白质将与其他遗传编码因子结合，开始赋予人工细胞一种基本的新陈代谢形式。此外，我们将利用已开发的途径来生产能够生产生物燃料的新型催化剂。铁-硫蛋白和铁-硫肽催化剂将在工业上得到潜在的应用。 我们是人工细胞合成领域的领导者之一。该项目将帮助我们巩固我们的地位，并有可能催生新技术和新公司。此外，该项目将丰富我们对生物学化学基础的理解。