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Directional multipart in vivo assembly and genome integration using Saccharomyces cerevisiae

Boekbijdrage - Boekabstract Conferentiebijdrage

Although various state-of-the-art metabolic engineering tools are by now well established for Saccharomyces cerevisiae, the introduction of multiple genes in its genome is still a laborious task. Yet, this is crucial for the stable expression of balanced biosynthesis pathways, i.e. for the efficient production of target molecules. To tackle this problem, we are developing a new technique called genomic Serine Integrase Recombinational Assembly (gSIRA). By exploiting the Streptomyces phiC31 integrase and its att sites as landing paths in the yeast’s genome, a whole pathway of genes can be directionally assembled and integrated at a desired locus. This is tested first with fluorescent proteins and will later on be evaluated for a small pathway of up to five genes. Benefits of this new assembly technique are 1) its time-saving nature (no vector intermediate needed), 2) its ability to manage multi-part assemblies and 3) the fact that it can be extended to introduce multiple copies of a pathway at various locations in the same single step. The only prerequisite is that one needs a library of baker’s yeast strains with the inducible integrase and specific att sites at specific genome locations. Thus, future research will also include the construction of a library of different att sites, to increase stability of the yeast strains using multiple landing paths. This technique can hence be used to quickly develop an industrially stable production strain of Saccharomyces cerevisiae.
Boek: Enabling Technologies for Eukaryotic Synthetic Biology, Symposium abstracts
Aantal pagina's: 1
Jaar van publicatie:2015