Advances in promoter engineering: Novel applications and predefined transcriptional control

Abstract

Synthetic biology continues to progress by relying on more robust tools for transcriptional control, of which promoters are the most fundamental component. Numerous studies have sought to characterize promoter function, determine principles to guide their engineering, and create promoters with stronger expression or tailored inducible control. In this review, we will summarize promoter architecture and highlight recent advances in the field, focusing on the novel applications of inducible promoter design and engineering towards metabolic engineering and cellular therapeutic development. Additionally, we will highlight how the expansion of new, machine learning techniques for modeling and engineering promoter sequences are enabling more accurate prediction of promoter characteristics.

Engineered Escherichia coli Nissle 1917 with urate oxidase and an oxygen-recycling system for hyperuricemia treatment

“…In order to further promote its practicality, additional work is needed. With the help of synthetic biology, we hope to further enhance the strain’s UA degradation rate through promoter engineering, 65 RBS engineering, 66 and expression timing. 67 , 68 The change of the order of temporal and spatial expression of genes in this artificial pathway could reduce the heterologous overexpression pressure to the cells and improve their ability to colonize the intestine.…”

Plant Flavonoid Production in Bacteria and Yeasts

“…However, such strategies largely depend on the specifications of the biosensors, thereby necessitating the reconstruction of biosensors for functional fine-tuning or improvement. To this end, both rational and evolutionary methodologies for engineering synthetic promoters (Snoek et al, 2020;Cazier and Blazeck, 2021;Tominaga et al, 2021) can be adapted. The assembly of well-optimized biosensors into genetic circuits to control the important pathway enzymes will further sophisticate the flavonoids production as well as balancing their expression levels via translational control using a ribosome binding site (RBS) library (Wang et al, 2021).…”

Genetic engineering for enhanced production of a novel alkaline protease BSP-1 in Bacillus amyloliquefaciens

“…In addition, by combining the transcriptional activator Spo0A with the regulatory region of Bacillus licheniformis , Zhou et al successfully achieved 1.46-fold increase in the yield of alkaline protease AprE ( Zhou et al, 2020 ). In addition, since promoter is one of the most critical factors in gene expression regulation ( Kumar and Bansal, 2018 ; Brázda et al, 2021 ; Cazier and Blazeck, 2021 ; Jensen and Galburt, 2021 ), its engineering also serves as an important strategy to increase microbial gene expression and metabolites production. Su et al substituted the wild-type −10 box and −35 box of aprN promoter, resulting in significantly higher nattokinase production than those previously reported in B. subtilis ( Wu et al, 2011 ).…”