Optimizing frameshift suppressor tRNAs for the recoding of the genetic code

有了这个奖项，化学部门的生命过程化学项目资助埃默里大学的Christine Dunham博士研究控制蛋白质表达的机制，目标是用不同的、非天然的化学成分来设计蛋白质。细胞的翻译机制负责将22种氨基酸整合到蛋白质中。从研究和应用的角度来看，非天然氨基酸的掺入是非常重要的，并且一直被认为是合成生物学的一个重要目标。这代表了当前的技术挑战。邓纳姆博士实验室的研究开发了一种解决方案，该方案基于一种名为转移RNA （tRNAs）的工程分子的使用。她的实验室的研究是研究生和博士后的训练基地，他们在生物学和化学的界面上获得跨学科的科学技能。该项目还被整合到一个外展计划中，该计划的使命是提高高中生对STEM的兴趣，培养这些学生成为独立的思考者，并帮助建立一支多元化的STEM劳动力队伍。如何操纵核糖体来表达含有一种或多种新化学成分的蛋白质是一个重要的合成生物学难题。认识到遗传密码不是不可改变的，重新编码的事件可能发生，吸引了许多化学生物学家的兴趣。先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性，以及由于与内源性trna的竞争，工程分子在核糖体上的结合率较低。本研究的重点是利用基于结构的移码抑制子trna的合理重新设计以及生物化学和结构生物学方法的新颖创新策略来扩展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界至关重要的话题。该奖项反映了美国国家科学基金会的法定使命，并通过使用基金会的知识价值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.