Bilateral BBSRC/NSF/BIO:Rewritable biocomputers in mammalian cells

双边 BBSRC/NSF/BIO：哺乳动物细胞中的可重写生物计算机

• 批准号: 1614642
• 负责人: Wilson Wong
• 金额: $ 66.47万
• 依托单位: Trustees of Boston University
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-08-15 至 2019-07-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1614642&HistoricalAwards=false
• 关键词: Bilateral; BBSRC; NSF; BIO; Rewritable

A major goal in engineering of biological systems is to predictably reprogram the genetics of cells to control a cellular function, such as to produce readily detectable indicators of metabolic states or environmental conditions. Although recent years have seen numerous advances towards this goal, the sophistication, performance and scale of mammalian genetic circuits trail their microbial counterparts. In this collaborative project, investigators from the US (Boston University) and the UK (University of Warwick) combine experimentation and computational modeling to develop a new genetic technology tool that will enable mammalian cells to respond to a signal to reprogram genetic circuits with memory capabilities. The memory feature is crucial to enable recordings of past events in complex environments where continuous monitoring is not feasible, which will be potentially useful for biopharmaceutical production. This project provides interdisciplinary training opportunities for high school, undergraduate, and graduate students in quantitative experimental methods and cell biology. This project will develop new tools for controlling mammalian cell gene expression using a class of DNA modifying enzymes called serine integrases. Serine integrases have advantages over the better characterized tyrosine recombinases due to the presence of recombinase directionality factors that bind to serine integrases to modify its conformation, permitting the reverse reaction to occur. To achieve this goal the PIs will: 1) perform detailed analyses of serine integrase (Bxb1, PhiC31, TP901-1 and TG1) activity and specificity in mammalian cells; 2) design, build and test inducible, rewritable memory switches for biopharmaceutical production; and 3) design, construct, and test a serine recombinase-based genetic circuit with reversible memory capabilities. This work will provide in-depth characterization of the specificity and toxicity of serine integrases and develop a novel decoder system for the reversible genetic circuit through integrated experimentation and computational modeling.This collaborative US/UK project is supported by the US National Science Foundation and the UK Biotechnology and Biological Sciences Research Council.

生物系统工程的一个主要目标是可预测地重新编程细胞的遗传学以控制细胞功能，例如产生易于检测的代谢状态或环境条件的指标。尽管近年来在实现这一目标方面取得了许多进展，但哺乳动物遗传电路的复杂性、性能和规模仍落后于微生物。在这个合作项目中，来自美国（波士顿大学）和英国（华威大学）的研究人员将实验和计算模型结合起来，开发一种新的遗传技术工具，使哺乳动物细胞能够响应信号，从而重新编程具有记忆能力的遗传电路。 记忆功能对于在无法连续监测的复杂环境中记录过去的事件至关重要，这对于生物制药生产可能有用。该项目为高中生、本科生和研究生提供定量实验方法和细胞生物学方面的跨学科培训机会。该项目将开发新工具，使用一类称为丝氨酸整合酶的 DNA 修饰酶来控制哺乳动物细胞基因表达。丝氨酸整合酶比更好表征的酪氨酸重组酶具有优势，因为重组酶方向性因子的存在与丝氨酸整合酶结合以改变其构象，从而允许逆反应发生。为了实现这一目标，PI 将： 1) 对哺乳动物细胞中的丝氨酸整合酶（Bxb1、PhiC31、TP901-1 和 TG1）活性和特异性进行详细分析； 2）设计、构建和测试用于生物制药生产的可诱导、可重写存储开关； 3) 设计、构建和测试具有可逆记忆能力的基于丝氨酸重组酶的遗传电路。 这项工作将深入描述丝氨酸整合酶的特异性和毒性，并通过综合实验和计算建模开发一种用于可逆遗传电路的新型解码器系统。这项美国/英国合作项目得到了美国国家科学基金会和英国生物技术和生物科学研究委员会的支持。