In vitro informational approach to biopolymer network dynamics

生物聚合物网络动力学的体外信息方法

• 批准号: 0750133
• 负责人: Vincent Noireaux
• 金额: $ 44万
• 依托单位: University of Minnesota-Twin Cities
• 依托单位国家: 美国
• 项目类别: Continuing Grant
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-09-15 至 2012-08-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=0750133&HistoricalAwards=false
• 关键词: vitro; informational; approach; biopolymer; network

In this proposal the PI will study the assembly of biopolymer networks in vitro. The PIs approach couples information flow and actin cytoskeleton structures formation and dynamics, and it provides perspectives in material science and biotechnology. E. coli cell-free expression system has been engineered with new properties that include: the control of expression from cloned genes; the use of three different RNA polymerases and three different transcription factors with their respective promoters/operators libraries; the fine tuning of transcript and protein degradation; and three formats of expression: batch, emulsion, and vesicle in continuous mode. This tool will allow the PI to study the dynamics of cytoskeleton related mechanisms with the complete chain of information, in the format and size of the cell as well as over extended period of time. In the first part of the project, characterization of this versatile expression system will be completed. A gene networks will be engineered to control quantitatively and accurately the expression of genes in time. This work will be used in the second part of the project to induce actin networks formation in a two dimensional unlimited space by expressing actin binding and polymerizing proteins. The relationship between expression, diffusion and structure of crosslinked networks of actin filaments will be studied. Finally, actin networks will be polymerized at the inner membrane of synthetic vesicles. Mechanisms of space symmetry breaking, protrusion formation and force generation will be investigated. The main intellectual merit of the project is the development of an innovative method to study quantitatively the information and physics properties of biopolymer networks assembly and dynamics. The research approach will provide quantitative insights into cell biology of actin structures. Part of the broader impact of this project includes the training of undergraduate and graduate students in biophysics. The students will have a hands-on experience in the laboratory. The experiments will range from standard cloning to the understanding of pattern formation and force generation in living cells. Students will be also involved in the development of models and simulations of gene circuits and diffusion mechanisms. This work will have an impact on a variety of scientific fields, in biological physics and bioengineering. The approach will also make significant contributions to synthetic, systems and cell biology.

在这个提议中，PI将研究生物聚合物网络在体外的组装。该方法将信息流与肌动蛋白细胞骨架结构的形成和动力学结合起来，为材料科学和生物技术提供了新的视角。大肠杆菌无细胞表达系统已被改造成具有新的特性，包括：控制克隆基因的表达；使用三种不同的RNA聚合酶和三种不同的转录因子及其各自的启动子/操作子文库；转录物和蛋白质降解的微调；三种表达方式：批量、乳化、囊泡连续模式。该工具将允许PI通过完整的信息链研究细胞骨架相关机制的动力学，包括细胞的格式和大小以及延长的时间。在项目的第一部分，将完成这个多功能表达系统的表征。一个基因网络将被设计来定量和准确地控制基因的表达。这项工作将用于项目的第二部分，通过表达肌动蛋白结合和聚合蛋白，在二维无限空间诱导肌动蛋白网络的形成。我们将研究肌动蛋白丝交联网络的表达、扩散和结构之间的关系。最后，肌动蛋白网络将在合成囊泡的内膜上聚合。研究空间对称破缺、突出形成和力产生的机制。该项目的主要智力优势是开发了一种创新的方法来定量研究生物聚合物网络组装和动力学的信息和物理特性。该研究方法将为肌动蛋白结构的细胞生物学提供定量的见解。这个项目更广泛的影响包括培养生物物理学的本科生和研究生。学生们将在实验室里有实际的经验。实验范围将从标准的克隆到对活细胞模式形成和力产生的理解。学生还将参与基因电路和扩散机制的模型和模拟的发展。这项工作将对生物物理学和生物工程等各个科学领域产生影响。该方法还将对合成、系统和细胞生物学做出重大贡献。