The Applications of Promoter-gene-Engineered Biosensors

Abstract

A promoter is a small region of a DNA sequence that responds to various transcription factors, which initiates a particular gene expression. The promoter-engineered biosensor can activate or repress gene expression through a transcription factor recognizing specific molecules, such as polyamine, sugars, lactams, amino acids, organic acids, or a redox molecule; however, there are few reported applications of promoter-enhanced biosensors. This review paper highlights the strategies of construction of promoter gene-engineered biosensors with human and bacteria genetic promoter arrays with regard to high-throughput screening (HTS) molecular drugs, the study of the membrane protein’s localization and nucleocytoplasmic shuttling mechanism of regulating factors, enzyme activity, detection of the toxicity of intermediate chemicals, and probing bacteria density to improve value-added product titer. These biosensors’ sensitivity and specificity can be further improved by the proposed approaches of Mn2+ and Mg2+ added random error-prone PCR that is a technique used to generate randomized genomic libraries and site-directed mutagenesis approach, which is applied for the construction of bacteria’s “mutant library”. This is expected to establish a flexible HTS platform (biosensor array) to large-scale screen transcription factor-acting drugs, reduce the toxicity of intermediate compounds, and construct a gene-dynamic regulatory system in “push and pull” mode, in order to effectively regulate the valuable medicinal product production. These proposed novel promoter-engineered biosensors aiding in synthetic genetic circuit construction will maximize the efficiency of the bio-synthesis of medicinal compounds, which will greatly promote the development of microbial metabolic engineering and biomedical science.

Future perspective: high-throughput construction of new ultrasensitive cytokine and virion liquid chips for high-throughput screening (HTS) of anti-inflammatory drugs or clinical diagnosis and treatment of inflammatory diseases

“…The newly constructed cytokine and virion liquid sensors have significant value for clinical diagnosis and treatment of infectious and inflammatory diseases (Figs. 5 and 6) [27,36,42]. Engineering Research Center of Ta r g e t e d a n d I n n o v a t i v e Therapeutics.…”

“…The applied electrochemical sensor is very sensitive for monitoring electron transition of IFNAR2/IL10RA/ACE2/Spike/GPCR-molecular drug interaction in a small area of the virion solution, which can be compared with the electrical signal of the negative control virions (wild-type virions without IFNAR2/IL10RA/ACE2/ Spike or GPCR incorporation) to gain electrical signaling of clinical diagnosis and to identify effective drugs (Fig. 6b) [2,9,10,21,[26][27][28][29][30][31][32].…”

“…Among the 1400 FDA-approved drugs, 475 (34%) drugs targeting 108 non-odorant GPCRs have been identified. Although there are about 70% known to have the ligand, 30% of the GPCRs are still without an identified ligand[20,21,[26][27][28][29][30][31]. Thus, the structure of un-characterized GPCRs…”

Transporter Engineering for Microbial Manufacturing

“…To date, a variety of biosensors have been developed for detection of polyamines, sugars, lactams, amino acids, organic acids, and redox molecules. [ 35 ] Recently, a useful method called the transcriptome‐assisted metabolite sensing was developed to discover native biosensors for target metabolites in microbial cells. [ 36 ] However, identifying the relevant biosensor for each compound of interest is challenging, thereby limiting the application of this strategy in transporter mining.…”

Transcriptional factor engineering in microbes for industrial biotechnology

“…Biosensor based on TFs capable of indicating the biosynthesis level of the target compound, will bring about many conveniences and high efficiency in searching for the most promising producer cells. 57 TFs are indispensable in the setup of an HTS because biosensor-based methods primarily rely upon TF to control the circuit output. 58,59 A TF specifically responding to L-serine was engineered into a biosensor capable of indicating the L-serine in a single cell of Corynebacterium glutamicum.…”