Advances in riboswitch‐based biosensor as food samples detection tool

Abstract

Food safety has always been a hot issue of social concern, and biosensing has been widely used in the field of food safety detection. Compared with traditional aptamer-based biosensors, aptamer-based riboswitch biosensing represents higher precision and programmability. A riboswitch is an elegant example of controlling gene expression, where the target is coupled to the aptamer domain, resulting in a conformational change in the downstream expression domain and determining the signal output. Riboswitch-based biosensing can be extensively applied to the portable real-time detection of food samples. The numerous key features of riboswitch-based biosensing emphasize their sustainability, renewable, and testing, which promises to transform engineering applications in the field of food safety. This review covers recent developments in riboswitch-based biosensors. The brief history, definition, and modular design (regulatory mode, reporter, and expression platform) of riboswitch-based biosensors are explained for better insight into the design and construction. We summarize recent advances in various riboswitch-based biosensors involving theophylline, malachite green, tetracycline, neomycin, fluoride, thrombin, naringenin, ciprofloxacin, and paromomycin, aiming to provide general guidance for the design of riboswitch-based biosensors. Finally, the challenges and prospects are also summarized as a way forward stratagem and signs of progress.

Broad-Spectrum Detection of Tetracyclines by Riboswitch-Based Cell-Free Expression Biosensing

“…The prepared riboswitch-based CFE biosensor allows activation by simply using human body temperature, which makes the CFE platform potentially more promising. 19 We sought to leverage the advantages of riboswitch-based biosensing to create a new approach for on-site monitoring TCs (including TC, OTC, CTC, and DC). The TC RNA aptamer screened by Berens et al 22 was introduced into the host RNA to create a tetracycline artificial riboswitch, which could control the expression of the downstream reporter gene green fluorescent protein (GFP).…”

“…Thavarajah et al successfully constructed the first riboswitch-based CFE biosensor based on a fluoride aptamer and realized environmental fluoride detection with a detection limit of 2 ppm. Moreover, the construction of riboswitch-based CFE biosensors can further break through the limitations of whole-cell biosensors such as genetically modified organism problems and the obstruction of cell membrane. ,, The riboswitch-based CFE biosensor links complex biological concepts with fast, intuitive readouts, allowing testers to quickly obtain experimental results (within tens of minutes). These techniques are simple enough for untrained individuals and at a manageable cost to enable detection at field sampling points without the need for complex instrumentation.…”

“…Lyophilized components of the CFE system can even be stored in tube-based sensors at room temperature for up to a year. The prepared riboswitch-based CFE biosensor allows activation by simply using human body temperature, which makes the CFE platform potentially more promising …”

“…The inherent biocontainment issues of whole-cell expression systems also make the biosensors difficult to commercialize. 19 The cell-free expression (CFE) system can provide an open, easily tuned translation environment, speed up the programming procedure, and be lyophilized and rehydrated as needed, which may enable a breakthrough in riboswitch-based biosensors. 19 Thavarajah et al 20 successfully constructed the first riboswitch-based CFE biosensor based on a fluoride aptamer and realized environmental fluoride detection with a detection limit of 2 ppm.…”

“…19 The cell-free expression (CFE) system can provide an open, easily tuned translation environment, speed up the programming procedure, and be lyophilized and rehydrated as needed, which may enable a breakthrough in riboswitch-based biosensors. 19 Thavarajah et al 20 successfully constructed the first riboswitch-based CFE biosensor based on a fluoride aptamer and realized environmental fluoride detection with a detection limit of 2 ppm. Moreover, the construction of riboswitch-based CFE biosensors can further break through the limitations of whole-cell biosensors such as genetically modified organism problems and the obstruction of cell membrane.…”