AutoBioTechA Versatile Biofoundry for Automated Strain Engineering

• Published in: ACS synthetic biology Vol. 13; no. 7; pp. 2227 - 2237
• Main Authors: Rosch, Tobias Michael, Tenhaef, Julia, Stoltmann, Tim, Redeker, Till, Kösters, Dominic, Hollmann, Niels, Krumbach, Karin, Wiechert, Wolfgang, Bott, Michael, Matamouros, Susana, Marienhagen, Jan, Noack, Stephan
• Format: Journal Article
• Language: English
• Published: United States American Chemical Society 19.07.2024
• Subjects: automation; CRISPR/Cas9; modular cloning; Escherichia coli; Corynebacterium glutamicum
• ISSN: 2161-5063; 2161-5063
• DOI: 10.1021/acssynbio.4c00298

The inevitable transition from petrochemical production processes to renewable alternatives has sparked the emergence of biofoundries in recent years. Manual engineering of microbes will not be sufficient to meet the ever-increasing demand for novel producer strains. Here we describe the AutoBioTech platform, a fully automated laboratory system with 14 devices to perform operations for strain construction without human interaction. Using modular workflows, this platform enables automated transformations of Escherichia coli with plasmids assembled via modular cloning. A CRISPR/Cas9 toolbox compatible with existing modular cloning frameworks allows automated and flexible genome editing of E. coli. In addition, novel workflows have been established for the fully automated transformation of the Gram-positive model organism Corynebacterium glutamicum by conjugation and electroporation, with the latter proving to be the more robust technique. Overall, the AutoBioTech platform excels at versatility due to the modularity of workflows and seamless transitions between modules. This will accelerate strain engineering of Gram-negative and Gram-positive bacteria.