Increasing Amino Acid Diversity in In Vitro Translation with Expanded Genetic Alphabets

该项目由数学和物理科学理事会化学部的生命过程化学计划和生物科学理事会分子和细胞生物科学部的分子遗传机制集群共同资助，有了这个奖项，化学部的生命过程化学计划正在资助韦斯特海默科学技术研究所（TWIST）的史蒂文·本纳博士在应用分子进化基金会内，扩大生物创造的蛋白质的力量。下一代生物技术将使用生命系统的组成部分来制造非天然的生物聚合物，包括含有非天然氨基酸的蛋白质。该奖项将发展这种生物技术，同时加深我们对蛋白质如何在活细胞中产生的理解。生物技术生产的蛋白质可用于各种产品，如药品，化妆品，工业制造和洗衣粉。添加非天然氨基酸可能会为探索具有天然蛋白质无法达到的特性的蛋白质开辟一个视野。本项目培养本科生掌握尖端技术开发的方法。本项目设计的实验将联合收割机化学合成与分子生物学、实验室进化和生物信息学相结合，以控制蛋白质体外合成的每一步。这包括开发新的方法，通过充电催化剂的定向进化将非天然氨基酸置于非天然转移RNA分子上，化学合成新的核酸类似物以管理RNA生物合成和翻译中的不忠实性，以及理解翻译系统如何利用辅助因子来确定信使RNA翻译的起点和终点。 这些与最近由X射线晶体学支持的核糖体模型相结合，在这个空间构建了合成生物学的复杂图像。

This project is jointly funded by the Chemistry of Life Processes Program in the Division of Chemistry in the Directorate of Mathematical and Physical Sciences and the Molecular Genetic Mechanisms Cluster in the Division of Molecular and Cellular Biosciences in the Directorate for Biological Sciences.With this award, the Chemistry of Life Processes Program in the Chemistry Division is funding Dr. Steven Benner of The Westheimer Institute for Science and Technology (TWIST) within the Foundation for Applied Molecular Evolution to expand the power of biologically created proteins. The next generation of biotechnology will use components of living systems to create unnatural biopolymers, including proteins that contain unnatural amino acids. This award will develop this biotechnology, at the same time as deepening our understanding of how proteins are made in living cells. Proteins produced by biotechnology find use in products as diverse as pharmaceuticals, cosmetics, industrial manufacture, and laundry detergents. Adding unnatural amino acids to these may open up for exploration a horizon of proteins having properties that natural proteins cannot reach. The project trains undergraduate students in methods of cutting-edge technology development.The experiments designed for this project combine chemical synthesis with molecular biology, laboratory evolution, and bioinformatics to gain control over each step in the in vitro synthesis of proteins. This includes developing new ways to put unnatural amino acids onto unnatural transfer RNA molecules by directed evolution of charging catalysts, chemically synthesizing new nucleic acid analogs to manage infidelity in RNA biosynthesis and translation, and understanding of how translation systems exploit auxiliary factors to determine the starting point and ending point of messenger RNA translation. These are coupled with models for the ribosome, recently supported by X-ray crystallography, to build a complex picture of synthetic biology in this space.