Bilateral BBSRC NSF/BIO - Synthetic gene circuits to measure and mitigate translational stress during heterologous protein expression

生物技术利用重组基因表达来生产一系列药物，包括胰岛素、疫苗和基于抗体的新型抗癌治疗剂。例如，人类胰岛素基因已被引入大肠杆菌，以推动这种有价值的药物在新的细菌宿主中廉价而安全地生产。在这项提案中，一个由生物学家和物理学家组成的跨学科团队将建立新的技术，以提高细胞以更高的效率和准确性制造重组蛋白的能力，从而提高下一代药物的质量、产量、成本效益和安全性。生物技术的大多数产品是蛋白质，一种被称为氨基酸的长链单位，其中有20种不同的品种。蛋白质的结构和氨基酸的序列是由基因决定的，核苷酸的DNA链具有特定的序列。为了制造蛋白质，基因内核苷酸序列中的编码信息首先被复制到信使RNA （mRNA）中，信使RNA也由核苷酸组成。然后，细胞中被称为核糖体的分子机器读取mRNA中的信息，产生正确的氨基酸链，形成蛋白质，这一过程被称为翻译。蛋白质中氨基酸的序列决定了它的性质和功能。当一个细胞被编程产生重组蛋白时，在翻译过程中，当一个氨基酸被选择添加到蛋白质中时，可能会发生错误。这些错误可以改变合成蛋白质的性质，并使其有缺陷；在一种蛋白质被用作药物的情况下，这可能会阻止有效的治疗，并且在最坏的情况下，对患者构成危险。在这项提案中，研究小组将与一家生物技术公司合作开发检测这类错误的敏感设备，并利用它们来了解细胞蛋白质制造机器如何以及何时出错，从而在未来将其最小化。然后，该团队将使用合成生物学方法中的基因组合来设计用于工业发酵罐的新型细胞，这些细胞能够在蛋白质产生时防止这些错误。我们将使用先进的数学模型来指导这些新的合成生物学基因电路的设计和安全性。总的来说，本提案中所描述的跨学科方法将涉及生物学家和物理学家共同努力，以创建改善新一代有效药物生产的系统。为了实现这些改进，它还将提供对细胞用于表达其基因和准确制造重组蛋白的基本机制的深入了解。更广泛地说，它将为优化一系列对生物技术和医学很重要的修饰蛋白的生产指明明确的路线。

Biotechnology uses recombinant gene expression to produce a range of medicines, including insulin, vaccines, and new anti-cancer therapeutic agents based upon antibodies. For instance, the human insulin gene has been introduced into the bacterium E. coli to drive the production of this valuable medicine in the new bacterial host, cheaply and safely. In this proposal, an interdisciplinary team of biologists and physicists will establish novel technologies to improve the ability of a cell to make recombinant proteins at higher efficiency, and with greater accuracy, improving the quality, yield, cost-effectiveness and safety of next-generation medicines. Most products of biotechnology are proteins, long chains of units called amino acids, of which there are 20 different varieties. The structures of proteins, and the sequence of their amino acids, are determined by genes, DNA strands of nucleotides with a specific sequence. To make a protein, the coding information locked in the sequence of nucleotides within the gene is first copied into a messenger RNA (mRNA), also composed of nucleotides. Then a molecular machine in the cell called a ribosome reads the information within the mRNA to produce the correct chain of amino acids, forming the protein, in a process called translation. The sequence of amino acids in the protein defines its properties and function. When a cell is programmed to produce a recombinant protein, errors can occur during translation, when an amino acid is selected to add to the protein. These errors can change the nature of the manufactured protein, and can make it defective; in the case of a protein being used as a medicine, this can prevent effective treatment, and as a worst case scenario, represent a danger to the patient.In this proposal, the research team will work with a biotechnology company to develop sensitive devices to detect this type of error, and use them to understand how and when the cellular protein manufacturing machinery makes mistakes, so they can be minimised in the future. The team will then use assemblies of genes in a synthetic biology approach to engineer new types of cells, designed to be used in industrial fermenters, that are capable of preventing these mistakes as the proteins are produced. We will use advanced mathematical models to guide the design and safety of these new synthetic biology gene circuits. Overall, the interdisciplinary approach described in this proposal will involve biologists and physicists working together to create systems that improve production of new generations of effective medicines. To allow these improvements, it will also provide insight into the fundamental mechanisms a cell uses to express its genes and make recombinant proteins accurately. More broadly, it will indicate clear routes to optimise production of a range of modified proteins important for biotechnology and medicine.