Fengjie Zhao, Christina M. Niman, Ghazaleh Ostovar, Marko S. Chavez, Joshua T. Atkinson, Benjamin M. Bonis, Jeffrey A. Gralnick, Mohamed Y. El-Naggar and James Q. Boedicker*,
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引用次数: 0
摘要
光遗传学是一种对基因表达进行时空控制的强大工具。目前已开发出多种光诱导基因调控因子在细菌中发挥作用,这些调控回路已被移植到新的宿主菌株中。在这里,我们开发了一种红光诱导转录因子,并将其应用于Shewanella oneidensis。该调控电路基于 iLight 光遗传系统,该系统利用红光控制基因表达。我们利用热力学模型和启动子工程改造了这一系统,使其在S. oneidensis宿主菌株中实现了光照和黑暗条件下的不同基因表达。我们进一步改进了 iLight 光遗传系统,添加了一个抑制因子,使基因回路反转,在红光照射下激活基因表达。倒置的 iLight 基因回路被用于控制 S. oneidensis 的胞外电子传递。同时使用红光和蓝光诱导的光遗传回路的能力也得到了证实。我们的工作拓展了 S. oneidensis 的合成生物学能力,这将促进未来电生细菌应用的进步。
Red-Light-Induced Genetic System for Control of Extracellular Electron Transfer
Optogenetics is a powerful tool for spatiotemporal control of gene expression. Several light-inducible gene regulators have been developed to function in bacteria, and these regulatory circuits have been ported to new host strains. Here, we developed and adapted a red-light-inducible transcription factor for Shewanella oneidensis. This regulatory circuit is based on the iLight optogenetic system, which controls gene expression using red light. A thermodynamic model and promoter engineering were used to adapt this system to achieve differential gene expression in light and dark conditions within a S. oneidensis host strain. We further improved the iLight optogenetic system by adding a repressor to invert the genetic circuit and activate gene expression under red light illumination. The inverted iLight genetic circuit was used to control extracellular electron transfer within S. oneidensis. The ability to use both red- and blue-light-induced optogenetic circuits simultaneously was also demonstrated. Our work expands the synthetic biology capabilities in S. oneidensis, which could facilitate future advances in applications with electrogenic bacteria.
期刊介绍:
The journal is particularly interested in studies on the design and synthesis of new genetic circuits and gene products; computational methods in the design of systems; and integrative applied approaches to understanding disease and metabolism.
Topics may include, but are not limited to:
Design and optimization of genetic systems
Genetic circuit design and their principles for their organization into programs
Computational methods to aid the design of genetic systems
Experimental methods to quantify genetic parts, circuits, and metabolic fluxes
Genetic parts libraries: their creation, analysis, and ontological representation
Protein engineering including computational design
Metabolic engineering and cellular manufacturing, including biomass conversion
Natural product access, engineering, and production
Creative and innovative applications of cellular programming
Medical applications, tissue engineering, and the programming of therapeutic cells
Minimal cell design and construction
Genomics and genome replacement strategies
Viral engineering
Automated and robotic assembly platforms for synthetic biology
DNA synthesis methodologies
Metagenomics and synthetic metagenomic analysis
Bioinformatics applied to gene discovery, chemoinformatics, and pathway construction
Gene optimization
Methods for genome-scale measurements of transcription and metabolomics
Systems biology and methods to integrate multiple data sources
in vitro and cell-free synthetic biology and molecular programming
Nucleic acid engineering.