Next-Generation Biological Biosensors for Trace Explosive Detection

Abstract

Survivors of the war experience lasting resident explosive left-over in the form of 2,4-dinitrotoluene (2,4-DNT), trinitrotoluene (TNT) that create enormous ecological and citizen safety challenges. Conventional approaches of detection are usually costly, time consuming or intrusive to nature. This paper introduces a gene expression-based biosensor constructed with the metabolically versatile soil bacterium Pseudomonas putida, a non-pathogenic, likely due to a gene expression-based biosensor built on a bacterium. Using controlled exposure experiments, it was found that the key genes PxylA, PaccD, and P3027 have reliably shown upregulation in reaction to 2,4-DNT during controlled exposure experiments. We have also used an engineer method by applying a CRISPR-Cas9 that allows the coupling of these gene promoters to reporter elements to obtain measurable or visible results when facing the detection of explosive residues. Dual UTR elements and synthetic promoter libraries were combined to balance the expression and provide a more sensitive solution in changing environmental circumstances. The sensitivity of the sensor was determined by qRT-PCR, fluorescence assays and viability tests over a series of DNT concentrations and in variant environmental conditions. The outcomes show strong signal induction, great specificity, and the ability to be used in the field. The study provides the next step in developing cost effective, deployable in field microbial biosensors and provides a principle framework of interaction with hybrid AI systems in the future of environmental detection.