{"database":"biostudies-literature","file_versions":[],"scores":null,"additional":{"omics_type":["Unknown"],"volume":["18(7)"],"submitter":["Mans R"],"pubmed_abstract":["Here, two methods are described for efficient genetic modification of Saccharomyces cerevisiae using CRISPR/Cas9. The first method enables the modification of a single genetic locus using in vivo assembly of a guide RNA (gRNA) expression plasmid without the need for prior cloning. A second method using in vitro assembled plasmids that could contain up to two gRNAs was used to simultaneously introduce up to six genetic modifications (e.g. six gene deletions) in a single transformation step by transforming up to three gRNA expression plasmids simultaneously. The method is not only suitable for gene deletion but is also applicable for in vivo site-directed mutagenesis and integration of multiple DNA fragments in a single locus. In all cases, the strain transformed with the gRNA expression plasmids was equipped with a genomic integration of Spcas9, leading to strong and constitutive expression of SpCas9. The protocols detailed here have been streamlined to be executed by virtually any yeast molecular geneticist."],"journal":["FEMS yeast research"],"full_dataset_link":["https://www.ebi.ac.uk/biostudies/studies/S-EPMC6074844"],"repository":["biostudies-literature"],"pubmed_title":["A protocol for introduction of multiple genetic modifications in Saccharomyces cerevisiae using CRISPR/Cas9."],"pmcid":["PMC6074844"],"pubmed_authors":["Wijsman M","Mans R","Daran-Lapujade P","Daran JM"],"additional_accession":[]},"is_claimable":false,"name":"A protocol for introduction of multiple genetic modifications in Saccharomyces cerevisiae using CRISPR/Cas9.","description":"Here, two methods are described for efficient genetic modification of Saccharomyces cerevisiae using CRISPR/Cas9. The first method enables the modification of a single genetic locus using in vivo assembly of a guide RNA (gRNA) expression plasmid without the need for prior cloning. A second method using in vitro assembled plasmids that could contain up to two gRNAs was used to simultaneously introduce up to six genetic modifications (e.g. six gene deletions) in a single transformation step by transforming up to three gRNA expression plasmids simultaneously. The method is not only suitable for gene deletion but is also applicable for in vivo site-directed mutagenesis and integration of multiple DNA fragments in a single locus. In all cases, the strain transformed with the gRNA expression plasmids was equipped with a genomic integration of Spcas9, leading to strong and constitutive expression of SpCas9. The protocols detailed here have been streamlined to be executed by virtually any yeast molecular geneticist.","dates":{"release":"2018-01-01T00:00:00Z","publication":"2018 Nov","modification":"2020-11-19T09:40:42Z","creation":"2019-03-26T23:50:30Z"},"accession":"S-EPMC6074844","cross_references":{"pubmed":["29860374"],"doi":["10.1093/femsyr/foy063"]}}