CAREER: Integrated Microsystems for Synthetic Biology

“这项奖励是根据2009年美国复苏和再投资法案（公法111-5）资助的。”这一建议旨在建立基础知识，使我们能够建立合成的多细胞机器。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、微制造的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利用化学微界面调节大肠杆菌合成信号通路的技术。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。Aim 2通过在微器件中添加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然后应用目标1和目标2中获得的知识，将控制从平面培养板转移到类似于简单多细胞系统的细胞球体组装上。这项工作的长期愿景是设计、构建和测试化学界面微系统，动态引导多细胞合成生物学系统的基因表达模式。这将需要开发用于感知和与细胞通信的相关微系统和对这些微系统作出反应的合成基因结构。今天的美国站在技术革命的边缘，就像我们在20世纪中期开始制造复杂的电子设备时所做的那样。本文介绍的工作采用微技术，从依赖同质细胞群的合成生物学标准方法中进行了彻底的范式转变。迄今为止，成功的合成模式生成只能通过手动将两种不同的细胞群（发送细胞和接收细胞）放置在细胞培养板上并允许它们进行通信来实现。这项工作旨在改变这一点：通过直接与程序化反应的细胞交流，可以使细胞培养在基因表达上产生空间不均匀性。创造复杂的基因表达空间模式的能力。合成细菌分化（Synthetic bacterial differentiation）将能够以一种类似于发育生物学中分化方案的方式，从合作细胞群体中构建出一类新的有机结构。广泛的影响广泛的影响和推广计划有五个组成部分：1)与Poplin出版社和Matthew Perry合作的K-12研讨会，开发和测试GoNano！2)根据PI发布的教学大纲为加州大学伯克利分校的研究生开设新的生物机械系统课程，以及为本科生开设新的核心MEMS课程（令人惊讶的是，加州大学伯克利分校的EECS中没有这门课程）；3)伯克利分校的领导参与？(4)与波多黎各大学马亚圭斯分校的Nelson Sepulveda教授进行学生交换和生物医学医学研讨会；(5)领导和参与伯克利的URM社区；PI曾担任密歇根大学拉丁裔教职员工协会主席，这种经验可以应用于伯克利。

"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.