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A cross-bacterial gene expression toolbox : engineering orthogonal transcription to enable predictable genetic engineering in diverse bacterial hosts

Cara Deal (UGent) , Daniel Ariza Calvo, Brecht De Paepe (UGent) , Antoni Planas, Lien De Wannemaeker (UGent) and Marjan De Mey (UGent)
(2024) 7th Applied Synthetic Biology in Europe (ASBE7), Abstracts.
Author
Organization
Project
Abstract
The ability to engineer predictability into biology is a core aim of synthetic biology. This is especially important in development of robust microbial cell factories that function in harsh industrial conditions. Genetic circuitry orthogonal to the host metabolism can help relieve environmental and host-related influences that often result in bioprocess variability, thus solving one of the main bottlenecks of industrial biotechnology. Existing orthogonal gene expression tools are limited in their host range and often exert significant metabolic burden on the host. This research aims to address these limitations by creating a gene expression toolbox that utilises the host’s native transcription machinery. Building on research showing that the sigma factor from one organism can control orthogonal gene expression in a second host, we aim to create a set of novel transcriptional control elements that function independently from environmental context in hosts across the bacterial domain, facilitating the transfer of genetic circuits between hosts of industrial interest. To this end, we demonstrate the use of rational library design and high-throughput screening methodologies to rewire the interaction between a sigma factor and its promoter. The result is a set of new-to-nature sigma factors that activate transcription at their cognate promoter sequence, without cross-talk to the genetic pathways involved in regulating host growth and development. The functionality of these transcriptional control elements is subsequently evaluated through testing in a range of bacterial hosts.

Citation

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MLA
Deal, Cara, et al. “A Cross-Bacterial Gene Expression Toolbox : Engineering Orthogonal Transcription to Enable Predictable Genetic Engineering in Diverse Bacterial Hosts.” 7th Applied Synthetic Biology in Europe (ASBE7), Abstracts, 2024.
APA
Deal, C., Ariza Calvo, D., De Paepe, B., Planas, A., De Wannemaeker, L., & De Mey, M. (2024). A cross-bacterial gene expression toolbox : engineering orthogonal transcription to enable predictable genetic engineering in diverse bacterial hosts. 7th Applied Synthetic Biology in Europe (ASBE7), Abstracts. Presented at the 7th Applied Synthetic Biology in Europe (ASBE7), Brno, Czech Republic.
Chicago author-date
Deal, Cara, Daniel Ariza Calvo, Brecht De Paepe, Antoni Planas, Lien De Wannemaeker, and Marjan De Mey. 2024. “A Cross-Bacterial Gene Expression Toolbox : Engineering Orthogonal Transcription to Enable Predictable Genetic Engineering in Diverse Bacterial Hosts.” In 7th Applied Synthetic Biology in Europe (ASBE7), Abstracts.
Chicago author-date (all authors)
Deal, Cara, Daniel Ariza Calvo, Brecht De Paepe, Antoni Planas, Lien De Wannemaeker, and Marjan De Mey. 2024. “A Cross-Bacterial Gene Expression Toolbox : Engineering Orthogonal Transcription to Enable Predictable Genetic Engineering in Diverse Bacterial Hosts.” In 7th Applied Synthetic Biology in Europe (ASBE7), Abstracts.
Vancouver
1.
Deal C, Ariza Calvo D, De Paepe B, Planas A, De Wannemaeker L, De Mey M. A cross-bacterial gene expression toolbox : engineering orthogonal transcription to enable predictable genetic engineering in diverse bacterial hosts. In: 7th Applied Synthetic Biology in Europe (ASBE7), Abstracts. 2024.
IEEE
[1]
C. Deal, D. Ariza Calvo, B. De Paepe, A. Planas, L. De Wannemaeker, and M. De Mey, “A cross-bacterial gene expression toolbox : engineering orthogonal transcription to enable predictable genetic engineering in diverse bacterial hosts,” in 7th Applied Synthetic Biology in Europe (ASBE7), Abstracts, Brno, Czech Republic, 2024.
@inproceedings{01JTGBAHB9B2335PJY9BHG52E0,
  abstract     = {{The ability to engineer predictability into biology is a core aim of synthetic biology. This is especially important in development of robust microbial cell factories that function in harsh industrial conditions. Genetic circuitry orthogonal to the host metabolism can help relieve environmental and host-related influences that often result in bioprocess variability, thus solving one of the main bottlenecks of industrial biotechnology.
Existing orthogonal gene expression tools are limited in their host range and often exert significant metabolic burden on the host. This research aims to address these limitations by creating a gene expression toolbox that utilises the host’s native transcription machinery. Building on research showing that the sigma factor from one organism can control orthogonal gene expression in a second host, we aim to create a set of novel transcriptional control elements that function independently from environmental context in hosts across the bacterial domain, facilitating the transfer of genetic circuits between hosts of industrial interest.
To this end, we demonstrate the use of rational library design and high-throughput screening methodologies to rewire the interaction between a sigma factor and its promoter. The result is a set of new-to-nature sigma factors that activate transcription at their cognate promoter sequence, without cross-talk to the genetic pathways involved in regulating host growth and development. The functionality of these transcriptional control elements is subsequently evaluated through testing in a range of bacterial hosts.}},
  author       = {{Deal, Cara and Ariza Calvo, Daniel and De Paepe, Brecht and Planas, Antoni and De Wannemaeker, Lien and De Mey, Marjan}},
  booktitle    = {{7th Applied Synthetic Biology in Europe (ASBE7), Abstracts}},
  language     = {{eng}},
  location     = {{Brno, Czech Republic}},
  title        = {{A cross-bacterial gene expression toolbox : engineering orthogonal transcription to enable predictable genetic engineering in diverse bacterial hosts}},
  year         = {{2024}},
}