Birth of a Photosynthetic Chassis: A MoClo Toolkit Enabling Synthetic Biology in the Microalga Chlamydomonas reinhardtii

Pierre Crozet (1) , Francisco Navarro (2) , Felix Willmund (3) , Payam Mehrshahi (2) , Kamil Bakowski (4) , Kyle Lauersen (5) , Maria-Esther Pérez-Pérez (6) , Pascaline Auroy (7, 8) , Aleix Gorchs Rovira (2) , Susana Sauret-Gueto (2) , Justus Niemeyer (3) , Benjamin Spaniol (3) , Jasmine Theis (3) , Raphael Trösch (3) , Lisa-Desiree Westrich (3) , Konstantinos Vavitsas (4) , Thomas Baier (5) , Wolfgang Hübner (5) , Felix de Carpentier (1) , Mathieu Cassarini (1) , Antoine Danon (1) , Julien Henri (1) , Christophe Marchand (1) , Marcello de Mia (1) , Kevin Sarkissian (1) , David Baulcombe (2) , G. Peltier (7, 8) , José-Luis Crespo (6) , Olaf Kruse (5) , Poul-Erik Jensen (4) , Michael Schroda (3) , Alison Smith (2) , Stéphane Lemaire (1)

1 IBPC (FR_550) - Institut de biologie physico-chimique
2 CAM - University of Cambridge [UK]
3 TU Kaiserslautern - Technische Universität Kaiserslautern
4 UCPH - University of Copenhagen = Københavns Universitet
5 Universität Bielefeld = Bielefeld University
6 Universidad de Sevilla = University of Seville
7 EBMP - Environnement, Bioénergie, Microalgues et Plantes
8 BIAM - Institut de Biosciences et Biotechnologies d'Aix-Marseille (ex-IBEB)

Microalgae are regarded as promising organisms to develop innovative concepts based on their photosynthetic capacity that offers more sustainable production than heterotrophic hosts. However, to realize their potential as green cell factories, a major challenge is to make microalgae easier to engineer. A promising approach for rapid and predictable genetic manipulation is to use standardized synthetic biology tools and workflows. To this end we have developed a Modular Cloning toolkit for the green microalga Chlamydomonas reinhardtii. It is based on Golden Gate cloning with standard syntax, and comprises 119 openly distributed genetic parts, most of which have been functionally validated in several strains. It contains promoters, UTRs, terminators, tags, reporters, antibiotic resistance genes, and introns cloned in various positions to allow maximum modularity. The toolkit enables rapid building of engineered cells for both fundamental research and algal biotechnology. This work will make Chlamydomonas the next chassis for sustainable synthetic biology.

Mots clés: modular cloning; Chlamydomonas reinhardtii; synthetic biology; algal biotechnology

Domaines: Biotechnologies; Botanique

Pierre Crozet, Francisco Navarro, Felix Willmund, Payam Mehrshahi, Kamil Bakowski, et al.. Birth of a Photosynthetic Chassis: A MoClo Toolkit Enabling Synthetic Biology in the Microalga Chlamydomonas reinhardtii. ACS Synthetic Biology, 2018, 7 (9), pp.2074-2086. ⟨10.1021/acssynbio.8b00251⟩. ⟨hal-01909673⟩