Tailoring baker's yeast Saccharomyces cerevisiae for functional testing of channelrhodopsin.

Channelrhodopsin 2 (ChR2) and its variants are the most frequent tools for remote manipulation of electrical properties in cells via light. Ongoing attempts try to enlarge their functional spectrum with respect to ion selectivity, light sensitivity and protein trafficking by mutations, protein engineering and environmental mining of ChR2 variants. A shortcoming in the required functional testing of large numbers of ChR2 variants is the lack of an easy screening system.

Novel auto-selection systems for transformation selection of Saccharomyces cerevisiae in rich complex media.

The most widely used strategy for selection of yeast transformed with episomal plasmids comprises the use of auxotrophic yeast strains in combination with vectors containing complementing prototrophic marker genes. Another approach uses heterologous genes or cassettes which, if present in the vector, render the otherwise sensitive yeast strain resistant to antibiotics. In addition, auto-selection systems for Saccharomyces cerevisiae have been developed that eliminate the requirement for synthetic drop-out media or the use of antibiotics for transformation selection and subsequent plasmid maintenance in expression cultures.

Trehalose as glucose surrogate in proliferation and cellular mobility of adult neural progenitor cells derived from mouse hippocampus.

The disaccharide trehalose (TRE) represents a natural energy supply for distinct non-mammalian species. Evidence has shown that TRE impacts on various properties including the stabilization of protein structure and cell membranes, which are important neuroprotective features against neurodegeneration. In this study, we tested the specific effect of TRE on cell proliferation and mobilization using an established experimental paradigm of adult neural progenitor cells (NPCs) derived from murine hippocampus.

Preloading budding yeast with all-in-one CRISPR/Cas9 vectors for easy and high-efficient genome editing.

The CRISPR/Cas9 technology has greatly improved genome editing in over recent years. However, several current CRISPR/Cas9 systems suffer from work-intensive cloning procedures and/or the requirement of co-transforming target cells with multiple system components simultaneously which can reduce the effectivity of such applications. Here, we present a new set of all-in-one CRISPR/Cas9 vectors that combine unique benefits of different already existent systems in order to further expand the technology's design possibilities.

Lipid determinants of endocytosis and exocytosis in budding yeast.

Cells maintain physicochemical characteristics of membranes in order to allow for proper function of membrane-associated cellular processes, such as endocytosis and exocytosis. To investigate the interplay between membrane properties and biological processes, we applied lipid engineering approaches that allowed for systematic manipulation of fatty acid unsaturation and sterol biosynthesis, the main regulators of membrane fluidity. In combination with electrophysiological membrane capacitance measurements, we were able to study the dependence of the endo- and exocytic activity of Saccharomyces cerevisiae on membrane lipid composition in vivo.