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: 2022
• 资助国家: 加拿大
• 起止时间: 2022-01-01 至 2023-12-31
• 项目状态: 已结题
• 来源: https://www.nserc-crsng.gc.ca/ase-oro/Details-Detailles_eng.asp?id=747995
• 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.

人造细胞是在实验室中用DNA、蛋白质和脂质构建的活细胞的仿制品。尽管在实验室中构建这样的系统比设计活细胞更具挑战性，但也有很多好处。首先，要理解生物学是如何工作的，没有比尝试从各个组成部分构建生物系统更好的方法了。此外，我们可以有意地加入在从头开始构建细胞模拟时有利的生物学部分，同时忽略生物学中有问题的部分。例如，非活细胞模拟物，即人造细胞，可以被构建成不复制的，这完全消除了对系统失去控制的恐惧。这不会造成长期的生态后果，因为一旦人造细胞分裂，环境就会恢复到原来的状态。我们已经制造出可以与自然活细胞进行化学交流和控制的人造细胞。这些技术有许多潜在的应用，从清洁环境中的污染物到治疗使人衰弱的疾病和感染。然而，最先进的人造细胞寿命并不长。也就是说，人造细胞只活跃几个小时，然后就会降解。在这里，我们打算赋予人造细胞代谢，使人造细胞能够长时间发挥作用。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一个更复杂部分的合成开始，即铁硫蛋白的合成。铁硫团簇是古老的无机原子排列，它们介导了维持细胞存活的大部分化学反应。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合成代谢铁硫蛋白的遗传编码途径。然后，这些蛋白质将与其他基因编码因子结合，开始赋予人造细胞一种基本的代谢形式。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。铁硫蛋白催化剂和铁硫肽催化剂都有可能在工业上得到应用。我们是人工细胞合成领域的领导者之一。这个项目将帮助我们巩固我们的地位，有可能产生新技术和新公司。此外，这个项目将丰富我们对生物学的化学基础的理解。