NDLI: Control of Gene Expression With Quercetin-Responsive Modular Circuits

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Control of Gene Expression With Quercetin-Responsive Modular Circuits

Content Provider	frontiers
Author	Kashiwagi, Fernanda Miyuki Wendler Miranda, Brenno de Oliveira Pedrosa, Fabio de Souza, Emanuel Maltempi Müller-Santos, Marcelo
Abstract	Control of gene expression is crucial for several biotechnological applications, especially for implementing predictable and controllable genetic circuits. Such circuits are often implemented with a transcriptional regulator activated by a specific signal. These regulators should work independently of the host machinery, with low gratuitous induction or crosstalk with host components. Moreover, the signal should also be orthogonal, recognized only by the regulator with minimal interference with the host operation. In this context, transcriptional regulators activated by plant metabolites as flavonoids emerge as candidates to control gene expression in bacteria. However, to engineer novel circuits requires the previous characterization of the genetic parts (e.g., genes, promoters, ribosome binding sites, and terminators) in the host of interest. Therefore, we decomposed the QdoR regulatory system of B. subtilis, responsive to the flavonoid quercetin, and reassembled its parts into genetic circuits programmed to have different levels of gene expression and noise dependent on the concentration of quercetin. We showed that only one of the promoters regulated by QdoR worked well in E. coli, enabling the construction of other circuits induced by quercetin. The QdoR expression was modulated with constitutive promoters of different transcriptional strengths, leading to low expression levels when QdoR was highly expressed and vice-versa. E. coli strains expressing high and low levels of QdoR were mixed and induced with the same quercetin concentration, resulting in two stable populations expressing different levels of their gene reporters. Besides, we demonstrated that the level of QdoR repression generated different noise levels in gene expression dependent on the concentration of quercetin. The circuits presented here can be exploited in applications requiring adjustment of gene expression and noise using a highly available and natural inducer as quercetin.
ISSN	22964185
DOI	10.3389/fbioe.2021.730967
Volume Number	9
Journal	Frontiers in Bioengineering and Biotechnology
Language	English
Publisher Date	2021-09-16
Access Restriction	Open
Subject Keyword	Quercetin Flavonoid E coli Genetic circuit QdoR
Content Type	Text
Resource Type	Article
Subject	Histology Bioengineering Biomedical Engineering Biotechnology

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