Cargando…

Control of Gene Expression With Quercetin-Responsive Modular Circuits

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...

Descripción completa

Detalles Bibliográficos
Autores principales: Kashiwagi, Fernanda Miyuki, Wendler Miranda, Brenno, de Oliveira Pedrosa, Fabio, de Souza, Emanuel Maltempi, Müller-Santos, Marcelo
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8481877/
https://www.ncbi.nlm.nih.gov/pubmed/34604189
http://dx.doi.org/10.3389/fbioe.2021.730967
_version_ 1784576779807096832
author Kashiwagi, Fernanda Miyuki
Wendler Miranda, Brenno
de Oliveira Pedrosa, Fabio
de Souza, Emanuel Maltempi
Müller-Santos, Marcelo
author_facet Kashiwagi, Fernanda Miyuki
Wendler Miranda, Brenno
de Oliveira Pedrosa, Fabio
de Souza, Emanuel Maltempi
Müller-Santos, Marcelo
author_sort Kashiwagi, Fernanda Miyuki
collection PubMed
description 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, engineering novel circuits requires the 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.
format Online
Article
Text
id pubmed-8481877
institution National Center for Biotechnology Information
language English
publishDate 2021
publisher Frontiers Media S.A.
record_format MEDLINE/PubMed
spelling pubmed-84818772021-10-01 Control of Gene Expression With Quercetin-Responsive Modular Circuits Kashiwagi, Fernanda Miyuki Wendler Miranda, Brenno de Oliveira Pedrosa, Fabio de Souza, Emanuel Maltempi Müller-Santos, Marcelo Front Bioeng Biotechnol Bioengineering and Biotechnology 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, engineering novel circuits requires the 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. Frontiers Media S.A. 2021-09-16 /pmc/articles/PMC8481877/ /pubmed/34604189 http://dx.doi.org/10.3389/fbioe.2021.730967 Text en Copyright © 2021 Kashiwagi, Wendler Miranda, de Oliveira Pedrosa, de Souza and Müller-Santos. https://creativecommons.org/licenses/by/4.0/This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
spellingShingle Bioengineering and Biotechnology
Kashiwagi, Fernanda Miyuki
Wendler Miranda, Brenno
de Oliveira Pedrosa, Fabio
de Souza, Emanuel Maltempi
Müller-Santos, Marcelo
Control of Gene Expression With Quercetin-Responsive Modular Circuits
title Control of Gene Expression With Quercetin-Responsive Modular Circuits
title_full Control of Gene Expression With Quercetin-Responsive Modular Circuits
title_fullStr Control of Gene Expression With Quercetin-Responsive Modular Circuits
title_full_unstemmed Control of Gene Expression With Quercetin-Responsive Modular Circuits
title_short Control of Gene Expression With Quercetin-Responsive Modular Circuits
title_sort control of gene expression with quercetin-responsive modular circuits
topic Bioengineering and Biotechnology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8481877/
https://www.ncbi.nlm.nih.gov/pubmed/34604189
http://dx.doi.org/10.3389/fbioe.2021.730967
work_keys_str_mv AT kashiwagifernandamiyuki controlofgeneexpressionwithquercetinresponsivemodularcircuits
AT wendlermirandabrenno controlofgeneexpressionwithquercetinresponsivemodularcircuits
AT deoliveirapedrosafabio controlofgeneexpressionwithquercetinresponsivemodularcircuits
AT desouzaemanuelmaltempi controlofgeneexpressionwithquercetinresponsivemodularcircuits
AT mullersantosmarcelo controlofgeneexpressionwithquercetinresponsivemodularcircuits