Modular, synthetic chromosomes as new tools for large scale engineering of metabolism
2022
Postma, Eline D. | Hassing, Else-Jasmijn | Mangkusaputra, Venda | Geelhoed, Jordi | De la Torre, Pilar | van den Broek, Marcel | Mooiman, Christiaan | Pabst, Martin | Daran, Jean-Marc | Daran-Lapujade, Pascale
The construction of powerful cell factories requires intensive genetic engineering for the addition of new functionalities and the remodeling of native pathways and processes. The present study demonstrates the feasibility of extensive genome reprogramming using modular, specialized de novo-assembled neochromosomes in yeast. The in vivo assembly of linear and circular neochromosomes, carrying 20 native and 21 heterologous genes, enabled the first de novo production in a microbial cell factory of anthocyanins, plant compounds with a broad range of pharmacological properties. Turned into exclusive expression platforms for heterologous and essential metabolic routes, the neochromosomes mimic native chromosomes regarding mitotic and genetic stability, copy number, harmlessness for the host and editability by CRISPR/Cas9. This study paves the way for future microbial cell factories with modular genomes in which core metabolic networks, localized on satellite, specialized neochromosomes can be swapped for alternative configurations and serve as landing pads for the addition of functionalities.
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