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生命过程化学项目和生物科学理事会分子和细胞生物科学部分子遗传机制集群共同资助。通过这个奖项，化学部门的生命过程化学项目资助了应用分子进化基金会韦斯特海默科学技术研究所（TWIST）的史蒂文·本纳博士，以扩大生物创造蛋白质的能力。下一代生物技术将使用生命系统的成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。这个奖项将发展这种生物技术，同时加深我们对活细胞中蛋白质是如何产生的理解。生物技术生产的蛋白质被广泛应用于药品、化妆品、工业制造和洗衣粉等产品中。将非天然氨基酸添加到这些蛋白质中，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新的领域。该项目培养本科生的尖端技术开发方法。本项目设计的实验将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。这包括通过充电催化剂的定向进化开发将非天然氨基酸放置到非天然转移RNA分子上的新方法，化学合成新的核酸类似物来管理RNA生物合成和翻译中的不确定性，以及理解翻译系统如何利用辅助因素来确定信使RNA翻译的起点和终点。这些与核糖体模型相结合，最近得到了x射线晶体学的支持，在这个领域建立了合成生物学的复杂图景。

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.