Genome optimization

CRISPR/Cas9-mediated genome editing

Graphic CRISPR/Cas9-mediated genome editing
Source: private

A fast and reliable Golden Gate-based cloning strategy, which allows us to target the Cas9 protein (molecular scissors) to multiple different locations throughout the genome, where it cuts double-stranded DNA to enable targeted insertions or deletions of DNA sequences. By using a nuclease deficient dCas9 protein, this system can also be used to up- or downregulate transcriptional levels of multiple genes in parallel.

Graphic CRISPR/Cas9-mediated genome editing
Source: private

Light-induced SCRaMbLE-ing of synthetic SC2.0 yeast genomes

Graphic Light-induced SCRaMbLE-ing
Source: private

Synthetic Sc2.0 chromosomes harbor symmetrical recombination sites (loxPsym) downstream of every non-essential gene. This enables to SCRaMbLE the whole genetic background in order to generate an optimized host organism for specific pathway engineering projects.

In our group, we established a red-light recombination system, dubbed L-SCRaMbLE, perfectly suited for the recombination and optimization of the yeast genome.

Graphic Light-induced SCRaMbLE-ing
Source: private

Hochrein L, Mitchell LA, Schulz K, Messerschmidt K, Mueller-Roeber B (2018) L-SCRaMbLE as a toolfor light-controlled Cre-mediatedrecombination in yeast. Nat Commun. 9: 1931. PMID: 29789561.