NO as a Sensor of Gene Expression Differences: Application to Synthetic Biology

NO 作为基因表达差异的传感器：在合成生物学中的应用

基本信息

批准号：
8398378
负责人：
Ryan Joseph Marcheschi
金额：
$ 5.39万
依托单位：
UNIVERSITY OF CALIFORNIA LOS ANGELES
依托单位国家：
美国
项目类别：
财政年份：
2012
资助国家：
美国
起止时间：
2012-01-01 至 2013-09-14
项目状态：
已结题

项目摘要

DESCRIPTION (provided by applicant): One of the primary goals of the fields of synthetic biology and metabolic engineering is to reprogram the behavior of organisms via the insertion of rationally engineered synthetic gene networks or the alteration of naturally occurring metabolic networks. A prominent component of this process is the need to balance gene expression to obtain optimal yields of desired products and avoid buildup of toxic intermediates. This proposal will address the current lack of methodology to serve this need by developing techniques for analyzing differences in dynamic gene expression and methods for accounting for and correcting those differences in the context of a metabolic pathway. The transient signaling molecule nitric oxide (NO) will be used for this purpose because it can be produced by NO synthases and degraded by NO dioxygenases or reductases, resulting in a tunable signal that will serve as an indicator of the difference in gene expression between two promoters. Coupled with a NO-sensitive promoter that will be developed as part of this research, the presence of NO will be used to drive gene expression of additional enzymes in response to an imbalance in the levels of NO production and degradation. As a test of this proposed system, I will apply it towards the production of 1-butanol (a potential biofuel) from threonine via the keto-acid biosynthesis metabolic pathway. In addition to the applications of this proposed research to synthetic biology and metabolic engineering, the development of a NO sensor for use in vivo will have applications to studies of bacterial antibiotic resistance and mammalian cell signaling and the development of a system to balance gene expression may have applications in the study of gene therapy.

描述（由申请人提供）：合成生物学和代谢工程领域的主要目标之一是通过插入合理工程的合成基因网络或自然发生的代谢网络的改变来重新编程生物体的行为。该过程的一个重要组成部分是需要平衡基因表达以获得所需产品的最佳产量，并避免堆积有毒中间体。该建议将通过开发用于分析动态基因表达的差异和在代谢途径背景下考虑和纠正这些差异的方法来解决目前缺乏来满足这种需求的方法。瞬态信号传导分子一氧化氮（NO）将用于此目的，因为它可以由NO合酶产生并被无二氧酶或还原酶降解，从而导致可调信号，这将是两个启动子之间基因表达的差异的指标。加上将作为这项研究的一部分开发的无敏感启动子，NO的存在将用于驱动其他酶的基因表达，以应对无生产和降解水平的不平衡。作为对该提出的系统的测试，我将通过酮酸生物合成代谢途径将其应用于从苏氨酸中生产1-丁醇（潜在的生物燃料）。除了这项拟议的研究在合成生物学和代谢工程中的应用外，在体内使用的无传感器的开发还将应用于细菌抗生素耐药性和哺乳动物细胞信号的研究，以及在基因治疗研究中的基因表达来平衡基因表达的系统的发展。