Birth of a photosynthetic chassis: a MoClo toolkit enabling synthetic biology in the microalga chlamydomonas reinhardtii

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.

This work was supported in part by Agence Nationale de la Recherche Grant ANR-17-CE05-0008 and LABEX DYNAMO ANR-LABX-011 (to P.C., A.D., F.d.C., J.H., C.H.M., M.d.M., K.S., M.C., and S.D.L.), by the DFG-funded TRR175 and FOR2092 (to F.W., B.S., J.N., J.T., L.W., R.T., and M.S.), by OpenPlant (BBSRC/EPSRC) (to F.J.N. and S.S.G.), by Ministerio de Economia y Competitividad Grants BFU2015-68216-P and BIO2015-74432-JIN (to J.L.C. and M.E.P.P.), by the VILLUM Foundation (Project no. 13363) (to P.E.J., K.B., K.V.), by the Technology Platforms at the Center for Biotechnology (CeBiTec) Bielefeld University (to K.L., T.B., and O.K.), by UK Biotechnology and Biological Sciences Research Council (BBSRC) (to P.M. and A.G.R.) and by ERA-SynBio project Sun2Chem (to P.A. and G.P.).