Beyond for modern lager brews: Exploring non- hybrids with heterotic maltotriose consumption and novel aroma profile.

Non-domesticated, wild yeasts have promising characteristics for beer diversification, particularly when used in the generation of interspecific hybrids. A major motivation for the current work was the question whether attractive novel interspecific hybrids can be created for the production of exotic lager beers without using the genomic resources of the ale yeast . Importantly, maltotriose utilization is an essential characteristic typically associated with domesticated ale/lager brewing strains.

Genome editing in Kluyveromyces and Ogataea yeasts using a broad-host-range Cas9/gRNA co-expression plasmid.

While CRISPR-Cas9-mediated genome editing has transformed yeast research, current plasmids and cassettes for Cas9 and guide-RNA expression are species specific. CRISPR tools that function in multiple yeast species could contribute to the intensifying research on non-conventional yeasts. A plasmid carrying a pangenomic origin of replication and two constitutive expression cassettes for Cas9 and ribozyme-flanked gRNAs was constructed.