Simplified CRISPR-Cas genome editing for Saccharomyces cerevisiae.
CRISPR-Cas has become a powerful technique for genetic engineering of yeast. Here, we present an improved version by using only one single plasmid expressing Cas9 and one or two guide-RNAs. A high gene deletion efficiency was achieved even with simultaneous recombination cloning of the plasmid and deletion in industrial strains.
Institute of Molecular Biosciences, Goethe University Frankfurt, 60438 Frankfurt am Main, Germany. Electronic address: generoso@bio.uni-frankfurt.de.
Wesley Cardoso
WC
Generoso
Institute of Molecular Biosciences, Goethe University Frankfurt, 60438 Frankfurt am Main, Germany. Electronic address: gottardi@bio.uni-frankfurt.de.
Manuela
M
Gottardi
Institute of Molecular Biosciences, Goethe University Frankfurt, 60438 Frankfurt am Main, Germany. Electronic address: m.oreb@bio.uni-frankfurt.de.
Mislav
M
Oreb
Institute of Molecular Biosciences, Goethe University Frankfurt, 60438 Frankfurt am Main, Germany. Electronic address: e.boles@bio.uni-frankfurt.de.
Eckhard
E
Boles
D064113:CRISPR-Cas Systems; D017353:Gene Deletion; D000072669:Gene Editing / Q000379:methods*; D005818:Genetic Engineering / Q000379:methods*; D016681:Genome, Fungal; D012441:Saccharomyces cerevisiae / Q000235:genetics*
CRISPR-Cas9; Metabolic engineering; Sacchromyces cerevisiae