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

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，例如产生代谢状态或环境条件的容易检测的指标。尽管近年来在实现这一目标方面取得了许多进展，但哺乳动物遗传回路的复杂性、性能和规模都落后于微生物。在这个合作项目中，来自美国（波士顿大学）和英国（沃里克大学）的研究人员将联合收割机实验和计算建模相结合，开发出一种新的遗传技术工具，使哺乳动物细胞能够对信号做出反应，重新编程具有记忆能力的遗传电路。 记忆功能对于在连续监测不可行的复杂环境中记录过去的事件至关重要，这对生物制药生产可能有用。该项目为高中，本科和研究生提供定量实验方法和细胞生物学的跨学科培训机会。该项目将开发使用一类称为丝氨酸整合酶的DNA修饰酶控制哺乳动物细胞基因表达的新工具。丝氨酸整合酶具有优于更好表征的酪氨酸重组酶的优势，这是由于重组酶方向性因子的存在，所述重组酶方向性因子结合丝氨酸整合酶以改变其构象，从而允许发生逆反应。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（Bxb 1、PhiC 31、TP 901 -1和TG 1）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、可重复记忆开关; 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。 这项工作将提供丝氨酸整合酶的特异性和毒性的深入表征，并通过综合实验和计算建模开发一种新的可逆遗传电路的解码器系统。这项美国/英国合作项目得到美国国家科学基金会和英国生物技术和生物科学研究理事会的支持。

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.