CAREER: Programming proteins by deep sequencing and design

职业：通过深度测序和设计对蛋白质进行编程

• 批准号: 1254238
• 负责人: Tim Whitehead
• 金额: $ 41.7万
• 依托单位: Michigan State University
• 依托单位国家: 美国
• 项目类别: Continuing Grant
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-05-15 至 2019-04-30
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1254238&HistoricalAwards=false
• 关键词: CAREER; Programming; proteins; deep; sequencing

1254238 WhiteheadProteins are central to a diverse set of fundamental activities necessary for cellular life. This same versatility leads to tremendous potential for proteins as designable agents in medicine such as monoclonal antibodies like Rituximab that can target and destroy non-Hodgkin lymphomas, and in industry such as enzymes that can deconstruct cellulosic biomass to simple carbohydrates. Yet natural proteins are not always optimal for industrial or medicinal applications, and current methods of improving proteins can be expensive and laborious. What is needed are new ways to search through the vast combinations of mutations to find the few that impart improvements to function. The primary research objective of this CAREER award is to design and test a massively parallel, rapid DNA deep sequencing method in order to quantitate sequence-function relationships for hundreds of thousands of protein variants. This method enables the rapid determination of those few beneficial mutations. The PI proposes to develop, apply, and integrate this method with computational design to reprogram specificity in natural and designed proteins. These optimized proteins will enable microbes to convert cellulosic biomass to next-generation fuels and value-added chemicals. The educational outreach of this CAREER award aims to increase the retention of women in science and engineering majors by creating additional opportunities for active participation in research. Towards this goal, the PI will integrate his educational and research objectives by: (i) utilizing existing MSU programs to place high school and undergraduate students in research projects in the PI's laboratory; (ii) teaching a series of workshops on biochemical engineering fundamentals for high school students; (iii) developing a new synthetic biology lab/lecture course at MSU for undergraduate students; and (iv) forming the first MSU team for the international genetically engineered machines (iGEM) competition, in which undergraduates perform self-guided research projects and present results at a national meeting. Women will be aggressively recruited to participate in each of these programs.This CAREER award is co-funded by the Division of Biological Infrastructure (DBI) through the Instrument Development for Biological Research (IDBR) program, and by the Division of Chemical, Bioengineering, Environmental, and Transport Systems (CBET).

1254238 Whitehead蛋白质是细胞生命所必需的各种基本活动的核心。同样的多功能性导致蛋白质在医学中作为可设计试剂的巨大潜力，例如单克隆抗体，如可以靶向和破坏非霍奇金淋巴瘤的利妥昔单抗，以及在工业中，例如可以将纤维素生物质解构为简单碳水化合物的酶。然而，天然蛋白质并不总是工业或医药应用的最佳选择，目前改进蛋白质的方法可能既昂贵又费力。我们需要的是新的方法来搜索大量的突变组合，以找到少数能改善功能的突变。 该奖项的主要研究目标是设计和测试一种大规模并行、快速的DNA深度测序方法，以定量数十万种蛋白质变体的序列-功能关系。这种方法能够快速确定那些少数有益的突变。PI建议开发，应用并整合这种方法与计算设计，以重新编程天然和设计蛋白质的特异性。这些优化的蛋白质将使微生物能够将纤维素生物质转化为下一代燃料和增值化学品。该职业奖的教育推广旨在通过创造更多积极参与研究的机会，增加科学和工程专业的女性保留率。为了实现这一目标，PI将通过以下方式整合其教育和研究目标：（i）利用现有的MSU计划，将高中和本科生放在PI实验室的研究项目中;（ii）为高中生讲授一系列生物化学工程基础知识的研讨会;（iii）为本科生开发MSU新的合成生物学实验室/讲座课程;（iv）为学生提供生物化学实验室课程。以及（iv）为国际基因工程机器（iGEM）竞赛组建第一支密歇根州立大学团队，在该竞赛中，本科生进行自我指导的研究项目，并在全国会议上展示结果。该CAREER奖由生物基础设施部（DBI）通过生物研究仪器开发（IDBR）计划以及化学、生物工程、环境和运输系统部（CBET）共同资助。