CAREER: Programming proteins by deep sequencing and design

1254238白头蛋白是细胞生命所必需的各种基本活动的中心。这种同样的多功能性使蛋白质在医学上作为可设计的试剂具有巨大的潜力，例如可以靶向并摧毁非霍奇金淋巴瘤的Rituximab等单抗，以及在工业中例如可以将纤维素生物量分解为简单碳水化合物的酶。然而，天然蛋白质在工业或医药应用中并不总是最佳的，而且目前改进蛋白质的方法可能昂贵且费力。现在需要的是一种新的方法来搜索大量的突变组合，以找到少数几个能改善功能的突变。该职业奖的主要研究目标是设计和测试一种大规模并行的快速DNA深度测序方法，以量化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为数不多的有益突变。PI建议开发、应用这种方法，并将其与计算设计相结合，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。这一职业奖的教育推广活动旨在通过创造更多的机会积极参与研究来留住科学和工程专业的女性。为了实现这一目标，该协会将整合他的教育和研究目标：(I)利用现有的密歇根州立大学项目，将高中生和本科生安排在其实验室的研究项目中；(Ii)为高中生教授一系列关于生化工程基础的工作坊；(Iii)在密歇根州立大学为本科生开发一个新的合成生物学实验室/讲座课程；以及(Iv)为国际基因工程机器(IGEM)竞赛组建第一个密歇根州立大学团队，由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。这一职业奖项由生物基础设施部(DBI)通过生物研究仪器开发(IDBR)计划和化学、生物工程、环境和运输系统司(CBET)共同资助。

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).