Design and construction of electrogenic cell-based biosensors for pathogens and toxins

病原体和毒素的基于细胞的生电生物传感器的设计和构建

• 批准号: BB/K016288/1
• 负责人: Martin Buck
• 金额: $ 60.91万
• 依托单位: Imperial College London
• 依托单位国家: 英国
• 项目类别: Research Grant
• 财政年份: 2013
• 资助国家: 英国
• 起止时间: 2013 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=BB%2FK016288%2F1
• 关键词: Design; construction; electrogenic; cell; based

The traditional laboratory-based analytical assays for bacterial pathogens and environmental toxins are expensive, time consuming and normally require specialised personnel and complex (expensive) equipment. This restricts their use in resource limited rural areas and developing countries where lack sufficient skilled personnel and healthcare facilities to rapidly identify the risks. There is therefore an urgent need to provide simple cost effective, fast on-site sensing solutions for pathogens and toxins associated with fatal bacterial infections and contaminated daily resources. For example, Vibrio cholerae bacteria contaminated water or food are typically responsible for the diarrheal illness cholera that can result in severe dehydration and even death within a matter of hours. Pseudomonas aeruginosa, commonly found on the surface of medical equipments such as catheters, is an opportunistic human pathogen and can be fatal for immunity compromised population. Arsenic contamination of groundwater is found in many places in the world, especially in Bangladesh where more than half of the population are facing the risk of arsenic poisoning which can cause cancers and various skin diseases. A simple, cost effective sensing solution that can accurately and rapidly report these severe health hazards will be vital to prevent the prevalence of these diseases and contribute to improved public health and quality of life.In this project we will establish a route to develop autonomous cell-based biosensors with direct digital electrical output for cost effective, on-site detection of pathogens and environmental toxins associated with fatal bacterial infectious diseases or contaminated resources. Different genetically coded sensors will be constructed and placed inside living benign gut bacteria Escherichia coli to monitor the appearance of specific toxin or fingerprinting molecules secreted by pathogen. The transduced sensory signals then further undergo amplification and modulation via customised genetic circuits to enhance sensing sensitivity and selectivity. Upon the detection of target pathogen or toxin, the genetically engineered bacterial cells will switch on the production of an electron shuttle to generate electrical power in a device known as microbial fuel cell, which utilises the modified bacteria to convert organic matter into electricity. As a result, autonomous cellular biosensors are constructed with the ability to power and report the associated pathogen or toxin levels to electronic systems without the use of specific assay equipment. The research builds a synthetic electron conduit between the currently less integrated cellular and electronic systems and will find many applications in environmental, agricultural and medical settings. The project will also contribute new design tools and methods, novel regulatory components and devices to the emerging field of deploying non-native biological systems in living microbes for repurposed actions of benefit to man.

传统的基于实验室的细菌病原体和环境毒素的分析测定是昂贵的、耗时的，并且通常需要专门的人员和复杂（昂贵）的设备。这限制了它们在资源有限的农村地区和发展中国家的使用，因为这些地区缺乏足够的熟练人员和医疗设施来快速识别风险。因此，迫切需要为与致命细菌感染和受污染的日常资源相关的病原体和毒素提供简单的成本有效的、快速的现场感测解决方案。例如，被霍乱弧菌污染的水或食物通常是导致霍乱的原因，霍乱可导致严重脱水，甚至在几小时内死亡。铜绿假单胞菌是一种常见于导管等医疗器械表面的条件致病菌，对免疫力低下人群具有致命性。世界上许多地方都发现地下水受到砷污染，特别是在孟加拉国，那里一半以上的人口面临砷中毒的风险，砷中毒可能导致癌症和各种皮肤病。一个简单、经济的传感解决方案，可以准确、快速地报告这些严重的健康危害，对于预防这些疾病的流行，改善公众健康和生活质量至关重要。在这个项目中，我们将建立一条路线，开发具有直接数字电输出的自主细胞生物传感器，现场检测与致命细菌性传染病或受污染资源有关的病原体和环境毒素。不同的基因编码传感器将被构建并放置在活的良性肠道细菌大肠杆菌中，以监测病原体分泌的特定毒素或指纹分子的出现。然后，转导的感觉信号通过定制的遗传电路进一步进行放大和调制，以增强传感灵敏度和选择性。一旦检测到目标病原体或毒素，基因工程细菌细胞将启动电子穿梭的生产，以在称为微生物燃料电池的设备中产生电力，该设备利用经修饰的细菌将有机物质转化为电力。因此，自主细胞生物传感器被构建为具有向电子系统供电和报告相关病原体或毒素水平的能力，而不使用特定的测定设备。该研究在目前集成度较低的蜂窝和电子系统之间建立了一个合成电子管道，并将在环境，农业和医疗环境中找到许多应用。该项目还将为在活微生物中部署非原生生物系统的新兴领域提供新的设计工具和方法，新颖的监管组件和设备，以重新调整对人类有益的行动。

项目成果

Amplification of small molecule-inducible gene expression via tuning of intracellular receptor densities.

• DOI: 10.1093/nar/gku1388
• 发表时间: 2015-02-18
• 期刊: Nucleic acids research
• 影响因子: 14.9
• 作者: Wang B;Barahona M;Buck M
• 通讯作者: Buck M

Rapid engineering of versatile molecular logic gates using heterologous genetic transcriptional modules.

• DOI: 10.1039/c4cc05264a
• 发表时间: 2014-10-11
• 期刊: Chemical communications (Cambridge, England)
• 作者: Wang B;Buck M
• 通讯作者: Buck M

Engineering modular and tunable genetic amplifiers for scaling transcriptional signals in cascaded gene networks.

• DOI: 10.1093/nar/gku593
• 发表时间: 2014-08
• 期刊: Nucleic acids research
• 影响因子: 14.9
• 作者: Wang B;Barahona M;Buck M
• 通讯作者: Buck M

Designer cell signal processing circuits for biotechnology.

• DOI: 10.1016/j.nbt.2014.12.009
• 发表时间: 2015-12-25
• 期刊: New biotechnology
• 影响因子: 5.4
• 作者: Bradley RW;Wang B
• 通讯作者: Wang B

Phenazines as model low-midpoint potential electron shuttles for photosynthetic bioelectrochemical systems.

作为光合生物电气化学系统的模型低中点电子班车的模型。

• DOI: 10.1039/d0sc05655c
• 发表时间: 2021-01-15
• 期刊: Chemical science
• 影响因子: 8.4
• 作者: Clifford ER;Bradley RW;Wey LT;Lawrence JM;Chen X;Howe CJ;Zhang JZ
• 通讯作者: Zhang JZ