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Proceedings of the National Academy of Sciences of the United States of America
11 03, 2020;117 (44): 27608-27619.

Synthetic gene-regulatory networks in the opportunistic human pathogen .

Robin A Sorg, Clement Gallay, Laurye Van Maele, Jean-Claude Sirard, Jan-Willem Veening
Author Information
  1. Robin A Sorg: Molecular Genetics Group, Groningen Biomolecular Sciences and Biotechnology Institute, Centre for Synthetic Biology, University of Groningen, 9747 AG, Groningen, The Netherlands.
  2. Clement Gallay: Department of Fundamental Microbiology, Faculty of Biology and Medicine, University of Lausanne, CH-1015 Lausanne, Switzerland. ORCID
  3. Laurye Van Maele: Université de Lille, CNRS, Inserm, Centre Hospitalier Universitaire de Lille, Institut Pasteur Lille, U1019-UMR 9017-CIIL-Center for Infection and Immunity of Lille, F-59000 Lille, France.
  4. Jean-Claude Sirard: Université de Lille, CNRS, Inserm, Centre Hospitalier Universitaire de Lille, Institut Pasteur Lille, U1019-UMR 9017-CIIL-Center for Infection and Immunity of Lille, F-59000 Lille, France. ORCID
  5. Jan-Willem Veening: Department of Fundamental Microbiology, Faculty of Biology and Medicine, University of Lausanne, CH-1015 Lausanne, Switzerland; jan-willem.veening@unil.ch. ORCID
PMID: 33087560 DOI: 10.1073/pnas.1920015117

Abstract

can cause disease in various human tissues and organs, including the ear, the brain, the blood, and the lung, and thus in highly diverse and dynamic environments. It is challenging to study how pneumococci control virulence factor expression, because cues of natural environments and the presence of an immune system are difficult to simulate in vitro. Here, we apply synthetic biology methods to reverse-engineer gene expression control in A selection platform is described that allows for straightforward identification of transcriptional regulatory elements out of combinatorial libraries. We present TetR- and LacI-regulated promoters that show expression ranges of four orders of magnitude. Based on these promoters, regulatory networks of higher complexity are assembled, such as logic AND gates and IMPLY gates. We demonstrate single-copy genome-integrated toggle switches that give rise to bimodal population distributions. The tools described here can be used to mimic complex expression patterns, such as the ones found for pneumococcal virulence factors. Indeed, we were able to rewire gene expression of the capsule operon, the main pneumococcal virulence factor, to be externally inducible (YES gate) or to act as an IMPLY gate (only expressed in absence of inducer). Importantly, we demonstrate that these synthetic gene-regulatory networks are functional in an influenza A virus superinfection murine model of pneumonia, paving the way for in vivo investigations of the importance of gene expression control on the pathogenicity of .

Keywords

Pneumococcus counterselection superinfection synthetic biology toggle switch

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MeSH Term

Animals
Bacterial Proteins
Disease Models, Animal
Gene Expression Regulation, Bacterial
Gene Regulatory Networks
Genes, Synthetic
Humans
Influenza A virus
Male
Mice
Nasopharynx
Operon
Opportunistic Infections
Pneumonia, Pneumococcal
Pneumonia, Viral
Promoter Regions, Genetic
Streptococcus pneumoniae
Superinfection
Synthetic Biology
Transcription Factors
Virulence Factors

Chemicals

Bacterial Proteins
Transcription Factors
Virulence Factors
Journal Article Research Support, Non-U.S. Gov't
Article has an altmetric score of 43

Word Cloud

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