Gene circuit engineering to improve the performance of a whole-cell lead biosensor

Jia, Xiaoqiang; Zhao, Tingting; Liu, Yilin; Bu, Rongrong; Wu, Kang

doi:10.1093/femsle/fny157

Cited by 28 publications

(22 citation statements)

References 37 publications

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“…Genetic amplifiers have been used to construct WCBs to intensify the output signal and increase sensitivity to analytes (22,23), but the effect of a positive feedback loop on increasing the sensitivity of the circuit is different from case to case, depending on the genetic context and the regulatory element it is coupled with. The LuxR positive feedback loop has been used for designing various WCBs, either increasing the output signal or improving the sensitivity, but has not been reported for detecting arsenic.…”

Section: Discussionmentioning

confidence: 99%

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Sensitive and Specific Whole-Cell Biosensor for Arsenic Detection

Jia

Zhao

et al. 2019

Appl Environ Microbiol

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Whole-cell biosensors (WCBs) have been designed to detect As(III), but most suffer from poor sensitivity and specificity. In this paper, we developed an arsenic WCB with a positive feedback amplifier in Escherichia coli DH5␣. The output signal from the reporter mCherry was significantly enhanced by the positive feedback amplifier. The sensitivity of the WCB with positive feedback is about 1 order of magnitude higher than that without positive feedback when evaluated using a halfsaturation As(III) concentration. The minimum detection limit for As(III) was reduced by 1 order of magnitude to 0.1 M, lower than the World Health Organization standard for the arsenic level in drinking water, 0.01 mg/liter or 0.13 M. Due to the amplification of the output signal, the WCB was able to give detectable signals within a shorter period, and a fast response is essential for in situ operations. Moreover, the WCB with the positive feedback amplifier showed exceptionally high specificity toward As(III) when compared with other metal ions. Collectively, the designed positive feedback amplifier WCB meets the requirements for As(III) detection with high sensitivity and specificity. This work also demonstrates the importance of genetic circuit engineering in designing WCBs, and the use of genetic positive feedback amplifiers is a good strategy to improve the performance of WCBs. IMPORTANCE Arsenic poisoning is a severe public health issue. Rapid and simple methods for the sensitive and specific monitoring of arsenic concentration in drinking water are needed. In this study, we designed an arsenic WCB with a positive feedback amplifier. It is highly sensitive and able to detect arsenic below the WHO limit level. In addition, it also significantly improves the specificity of the biosensor toward arsenic, giving a signal that is about 10 to 20 times stronger in response to As(III) than to other metals. This work not only provides simple but effective arsenic biosensors but also demonstrates the importance of genetic engineering, particularly the use of positive feedback amplifiers, in designing WCBs.A rsenic (As)-contaminated groundwater, occurring from mining or agriculture or natural contamination due to the abundance of arsenic in the Earth's crust, is a serious global health issue. Long-term exposure to arsenic can result in various diseases including cancers (1, 2). It is estimated that over 100 million people worldwide may be at risk from consuming water contaminated with arsenic (3). The World Health Organization (WHO) has recommended 0.01 mg/liter (0.13 M) as a safe permissible level for arsenic in drinking water (4), and the Food and Agriculture Organization (FAO) has set a maximum contamination level (MCL) for arsenic of 0.01 mg/liter in irrigation water (5).

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Section: Discussionmentioning

confidence: 99%

“…Genetic circuit engineering was done to improve the performance of the WCB. Positive feedback is common in nature and well known for signal amplification (22). It has been used to improve the sensitivity of WCBs in response to various analytes, including antibiotics, amino acids, and heavy metals (23).…”

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confidence: 99%

Sensitive and Specific Whole-Cell Biosensor for Arsenic Detection

Jia

Zhao

et al. 2019

Appl Environ Microbiol

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“…The e ciency of the predicted model design was con rmed experimentally, and the model may further be useful for preparing detailed modelling of not only regarding the Ar sensor, but also other types of bioreporters (Berset et al 2017). Jia et al (2018) introduced six gene circuits for the whole-cell-based biosensor for Pb by rearranging various regulatory elements in the Pb resistance operon and additionally incorporating positive feedback loops. The operon resistant for Pb encodes six genes and consists of regulatory pbrRT on one side and pbrABCD on the other side.…”

Section: Critical Parameters Used In Synthetic Biology: Biocomputation Aspectmentioning

confidence: 94%

In-situ monitoring of xenobiotics using genetically engineered whole-cell-based microbial biosensors: recent advances and outlook

Ali

Mittal

Kaur

2021

World J Microbiol Biotechnol

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Industrialisation, directly or indirectly, exposes humans to various xenobiotics. The increased magnitude of chemical pesticides and toxic heavy metals in the environment, as well as their intrusion into the food chain, seriously threatens human health. Therefore, the surveillance of xenobiotics is crucial for social safety and security. Online investigation by traditional methods is not su cient for the detection and identi cation of such compounds because of the high costs and their complexity. Advancement in the eld of genetic engineering provides a potential opportunity to use genetically modi ed microorganisms. In this regard, whole-cell-based microbial biosensors (WCBMB) represent an essential tool that couples genetically engineered organisms with an operator/promoter derived from a heavy metal-resistant operon combined with a regulatory protein in the gene circuit. The plasmid controls the expression of the reporter gene, such as gfp, luc, lux and lacZ, to an inducible gene promoter and has been widely applied to assay toxicity and bioavailability. This review summarises the recent trends in the development and application of microbial biosensors and the use of mobile genes for biomedical and environmental safety concerns.

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“…Genetic logic gates can also function as a biological filter and an amplifier ( Kim et al, 2016 ; Jia et al, 2018 , 2019 ; Saltepe et al, 2018 ). A positive feedback amplifier using LuxR auto tuning elements improved the sensitivity and specificity of an arsenic whole-cell bioreporter.…”

Section: Whole-cell Microbial Bioreporter Designs For Contaminant Meamentioning

confidence: 99%

“…The minimum detection limit was reduced by one order of magnitude. Similarly, a lead whole-cell bioreporter containing positive feedback amplifiers was 15–20 times more sensitive than the one without amplifiers ( Jia et al, 2018 ). Compared with the constitutively expressed regulator, the positive feedback regulator enhances the specificity and sensitivity of bioreporters ( Jia et al, 2019 ).…”

Section: Whole-cell Microbial Bioreporter Designs For Contaminant Meamentioning

confidence: 99%

Whole-Cell Microbial Bioreporter for Soil Contaminants Detection

Zeng

Chen

et al. 2021

Front. Bioeng. Biotechnol.

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Anthropogenic activities have released various contaminants into soil that pose a serious threat to the ecosystem and human well-being. Compared to conventional analytical methodologies, microbial cell-based bioreporters are offering a flexible, rapid, and cost-effective strategy to assess the environmental risks. This review aims to summarize the recent progress in the application of bioreporters in soil contamination detection and provide insight into the challenges and current strategies. The biosensing principles and genetic circuit engineering are introduced. Developments of bioreporters to detect and quantify heavy metal and organic contaminants in soil are reviewed. Moreover, future opportunities of whole-cell bioreporters for soil contamination monitoring are discussed.

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Gene circuit engineering to improve the performance of a whole-cell lead biosensor

Cited by 28 publications

References 37 publications

Sensitive and Specific Whole-Cell Biosensor for Arsenic Detection

Sensitive and Specific Whole-Cell Biosensor for Arsenic Detection

In-situ monitoring of xenobiotics using genetically engineered whole-cell-based microbial biosensors: recent advances and outlook

Whole-Cell Microbial Bioreporter for Soil Contaminants Detection

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