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

人造细胞是在实验室里用DNA、蛋白质和脂类建造的活细胞的模仿物。尽管在实验室建造这样的系统比设计活细胞更具挑战性，但它有很多好处。首先，可以说，没有比试图用组成部分构建生物系统更好的方式来理解生物是如何工作的。此外，当我们从头开始构建细胞模拟时，我们可以有意识地加入生物学中有利的部分，同时省略生物学中有问题的部分。例如，无生命的细胞模拟物，即人造细胞，可以被构建为不复制，这完全消除了对失去对系统的控制的恐惧。不可能有长期的生态后果，因为一旦人造细胞解体，环境就会恢复到以前的状态。我们已经建造了人造细胞，可以与自然活细胞进行化学交流并控制它们。这种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令人衰弱的疾病和感染。然而，人工细胞中最先进的技术并不是很持久。也就是说，人造细胞在几个小时内处于活动状态，然后降解。在这里，我们打算让人造细胞进行新陈代谢，以便人造细胞能够发挥足够长的功能，从而发挥作用。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。我们将从新陈代谢中更复杂的部分之一的合成开始，这是铁硫蛋白的合成。铁-硫团簇是古老的、无机的原子排列，参与维持细胞存活的大部分化学物质。通过利用我们在合成铁-硫簇和体外遗传学方面的专业知识，我们将在人造细胞内开发一种遗传编码的途径，合成代谢中的铁-硫蛋白。然后，这些蛋白质将与其他基因编码的因子结合，开始给人造细胞一种基本的新陈代谢形式。此外，我们将利用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在工业上具有潜在的应用前景。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，可能会催生新技术和新公司。此外，这个项目将丰富我们对生物学的化学基础的理解。

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.