CAREER: Integrated Microsystems for Synthetic Biology

“该奖项是根据2009年美国复苏和再投资法案（公法111-5）资助的。“这项提案旨在建立基础知识，使我们能够构建合成多细胞机器。研究人员将设计、制造和测试一系列混合系统，包括转基因细菌、微制造化学致动器阵列和光学传感。这项技术工作的主要目标是开发调节大肠杆菌中合成信号通路的技术。大肠杆菌细菌的化学微界面。目标1将产生一个微系统和一个合成基因结构，其中细胞在微系统的引导下，将表达由用户编程的基因表达模式。目标2通过在微器件中加入协调反馈来扩展这一点，以开发一个完整的混合双组分图灵反应扩散系统。目标3应用目标1和目标2中获得的知识，将控制从平板培养板过渡到类似于简单多细胞系统的球状细胞组装体。这项工作的长期愿景是设计、构建和测试化学界面微系统，以动态引导多细胞合成生物学系统的基因表达模式。这将需要开发用于传感和与细胞通信的相关微系统以及响应这些微系统的合成基因结构。今天的美国站在技术革命的边缘，就像我们在世纪中期开始制造复杂的电子设备时一样。这里介绍的工作采用微技术，使一个根本的范式转变，从标准的合成生物学方法，依赖于同质群体的细胞。迄今为止，成功的合成模式生成仅通过手动将两个不同的细胞群体（发送者和接收者细胞）放置在细胞培养板上并允许它们通信来实现。这项工作旨在改变这一点：通过与编程响应的细胞直接通信，可以使细胞培养物在基因表达中产生空间不均匀性。创造基因表达的复杂空间模式的能力，（即。合成细菌分化）将使得能够以类似于发育生物学中的分化方案的方式从协同操作的细胞群体构建一类新的有机构建体。广泛的影响和推广计划有五个组成部分：1）K-12讲习班与波普林出版社和马修佩里开发和测试GoNano！，一个基于卡片的教学辅助小学和初中学生，2）在加州大学伯克利分校的研究生的一个新的BioMEMS课程的发展，根据教学大纲上的PI已经出版，和一个新的核心MEMS课程的本科生（这，令人惊讶的是，不存在于EECS在加州大学伯克利分校），3）领导参与伯克利分校？的SUPERB计划，其中PI是校友，4）学生交流和生物MEMS研讨会与纳尔逊塞普尔韦达教授在波多黎各，马亚圭斯大学，5）领导和参与URM社区在伯克利分校; PI担任拉丁美洲教职员工协会在密歇根大学的主席，经验可以在伯克利分校应用。

"This award is funded under the American Recovery and Reinvestment Act of 2009 (Public Law 111-5)."This proposal aims at establishing the foundational knowledge that will enable us to build synthetic multi-cellular machines. The researchers will design, fabricate and test a series of hybrid systems consisting of genetically modified bacteria, microfabricated arrays of chemical actuators and optical sensing. The broad goal of the technical work is to develop techniques for modulating synthetic signaling pathways in E. coli bacteria using chemical micro-interfaces. Aim 1 will produce a microsystem and a synthetic gene construct in which cells, guided by the microsystem, will express gene expression patterns programmed by the user. Aim 2 expands this by adding coordinated feedback into the microdevices to develop a complete hybrid two-component Turing reaction-diffusion system. Aim 3 then applies the knowledge gained in Aims 1 and 2 to transition the control from a flat culture plate onto a spheroidal assembly of cells resembling a simple multi-cellular system.The long term vision of this work is to design, build and test chemical interface microsystems that dynamically guide gene expression patterns of multi-cellular synthetic biology systems. This will require the development of both the relevant microsystems intended for sensing and communicating with cells and the synthetic gene constructs which respond to these microsystems. The United States today stands on the brink of a technological revolution, much as we did in the mid-20th century when we began to build complex electronic devices. Intellectual MeritThe work presented here employs microtechnology to make a radical paradigm shift away from the standard methods in synthetic biology which rely on homogenous populations of cells. To date, successful synthetic pattern generation has only been achieved by manually placing two different populations of cells on a cell culture plate (sender and receiver cells) and allowing them to communicate. This work intends to change this: by communicating directly with cells programmed to respond, cultures of cells can be made to develop spatial inhomogeneities in gene expression. The ability to create complex spatial patterns of gene expression, (ie. synthetic bacterial differentiation) will enable the fabrication of a new class of organic constructs built from co-operating cell populations in a manner analogous to differentiation schemes in developmental biology. Broad ImpactThe broad impact and outreach plan has five components: 1) K-12 workshops with Poplin Press and Matthew Perry to develop and test GoNano!, a card-based teaching aid for elementary and junior high students, 2) the development of both a new BioMEMS course for graduate students at UC Berkeley, based on a syllabus on which the PI has published, and a new core MEMS course for undergraduates (which, surprisingly, does not exist in EECS at UC Berkeley), 3) leadership involvement in Berkeley?s SUPERB program, of which the PI is an alumnus, 4) student exchange and BioMEMS seminars with Prof. Nelson Sepulveda at the University of Puerto Rico, Mayaguez, 5) leadership and participation in the URM community at Berkeley; the PI served as the president of the Latino Faculty & Staff Association at the University of Michigan and that experience can be applied at Berkeley.