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Synthetic biology parts are transferable across organisms, enabling predictable gene expression. This study quantifies part behavior in multiple hosts, creating a lookup table for cross-organism engineering.

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Area of Science:

  • Synthetic Biology
  • Molecular Biology
  • Genetics

Background:

  • Developing new biological parts for each host organism is impractical.
  • Gene expression parts are known to be transferable, but quantitative data on this transferability is limited.

Purpose of the Study:

  • To systematically quantify the behavior of a standardized set of synthetic biology parts across multiple host organisms.
  • To establish a method for generalizing modular parts sets beyond a single host species.

Main Methods:

  • Developed a broad host range (BHR) plasmid system (openCIDAR) compatible with the CIDAR parts collection.
  • Tested DNA constructs across four Pseudomonadota species: Escherichia coli, Pseudomonas putida, Cupriavidus necator, and Komagataeibacter nataicola.
  • Quantified part performance using Molecules of Equivalent Fluorescein (MEFL) and applied linear regression to identify divergent parts.

Main Results:

  • The CIDAR parts enabled graded gene expression across all tested organisms, with similar expression trends but varying average levels.
  • A lookup table is required to translate designs between hosts to achieve consistent MEFL.
  • The promoter J23100 showed significantly different behavior in Komagataeibacter nataicola compared to other hosts.

Conclusions:

  • The openCIDAR system allows evaluation of CIDAR-compatible parts in multiple Proteobacteria, generalizing synthetic biology parts sets.
  • This approach accelerates engineering efforts in diverse species for various applications.
  • A few standardized parts sets could potentially span across the tree of life.