Optimizing frameshift suppressor tRNAs for the recoding of the genetic code

通过该奖项，化学部生命过程化学计划资助来自埃默里大学的Christine Dunham博士研究控制蛋白质表达的机制，目标是设计具有多种非天然化学部分的蛋白质。细胞的翻译机制负责蛋白质中22种氨基酸的结合。为了研究和应用的目的，非天然氨基酸的掺入是非常重要的，长期以来一直被认为是合成生物学的重要目标。这代表着当前的技术挑战。邓纳姆博士实验室的研究开发了一种基于被称为转移RNA(TRNAs)的工程分子的解决方案。她实验室的研究是研究生和博士后研究员的训练场，他们在生物和化学之间获得跨学科的科学技能。该项目还被纳入一个推广计划，其使命是提高高中生对STEM的兴趣，培养这些学生成为独立思考者，并帮助创建多样化的STEM工作队伍。如何操纵核糖体表达包含一个或多个新化学部分的蛋白质的问题是一个重要的合成生物学难题。遗传密码并不是一成不变的，重新编码事件可能会发生，这一认识吸引了许多化学生物学家的兴趣。以前解决这一难题的方法的主要局限性包括与宿主翻译机制的交叉反应，以及由于与内源tRNA的竞争而导致核糖体上工程分子的低掺入。这一建议的核心是利用合理的、基于结构的重新设计框架移位抑制因子tRNA以及生化和结构生物学方法的新颖和创新策略来扩大tRNA的编码能力。这项研究的信息为蛋白质合成的调控提供了新的见解，这是一个对广泛的生物分子和合成生物学研究社区至关重要的话题。这一奖项反映了NSF的法定使命，并通过使用基金会的智力优势和更广泛的影响审查标准进行评估，被认为值得支持。

With this award, the Chemistry of Life Processes Program in the Chemistry Division is funding Dr. Christine Dunham from Emory University to investigate mechanisms to control protein expression with the goal of engineering proteins with diverse, non-natural chemical moieties. The translational machinery of cells is responsible for the incorporation in proteins of twenty-two amino acids. For research and application purposes, the incorporation of non-naturally occurring amino acids is very important and has long been recognized as an important goal of synthetic biology. This represents a current technological challenge. The research in Dr. Dunham's lab develops a solution to this challenge that is based on the use of engineered molecules called transfer RNA (tRNAs). The research in her lab is a training ground for graduate students and postdoctoral fellows, who acquire interdisciplinary scientific skills at the interface between biology and chemistry. This project is also integrated into an outreach program whose mission is to raise the interest of high school students in STEM, to train these students to become independent thinkers, and to help create a diverse STEM workforce.The question of how to manipulate the ribosome to express proteins containing one or more novel chemical moieties is an important synthetic biology puzzle. The realization that the genetic code is not immutable and that recoding events could occur has attracted the interest of many chemical biologists. The major limitations of previous approaches to solve this puzzle include cross-reactivity with host translation machinery and low incorporation of engineered molecules on the ribosome due to competition with endogenous tRNAs. This proposal centers on expanding the coding capacity of tRNAs using the novel and innovative strategy of rational, structure-based redesign of frameshift suppressor tRNAs and biochemical and structural biology approaches. Information from this study provides new insights into the regulation of protein synthesis, a topic critically important to a broad biomolecular and synthetic biology research community.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.