title: The application of the Cre-loxP system for generating multiple knock-out and knock-in targeted loci.
abstract: Dictyostelium discoideum is an exceptionally powerful eukaryotic model to study many aspects of growth, development, and fundamental cellular processes. Its small-sized, haploid genome allows highly efficient targeted homologous recombination for gene disruption and knock-in epitope tagging. We previously described a robust system for the generation of multiple gene mutations in Dictyostelium by recycling the Blasticidin S selectable marker after transient expression of the Cre recombinase. We have now further optimized the system for higher efficiency and, additionally, coupled it to both, knock-out and knock-in gene targeting, allowing the characterization of multiple and cooperative gene functions in a single cell line.
authors.affiliation: Institute for Biophysical Chemistry, Hannover Medical School, Hannover, Germany. Faix.Jan@mh-hannover.de
authors.forename: Jan
authors.initials: J
authors.lastname: Faix
authors.forename: Joern
authors.initials: J
authors.lastname: Linkner
authors.forename: Benjamin
authors.initials: B
authors.lastname: Nordholz
authors.forename: James L
authors.initials: JL
authors.lastname: Platt
authors.forename: Xin-Hua
authors.initials: XH
authors.lastname: Liao
authors.forename: Alan R
authors.initials: AR
authors.lastname: Kimmel
mesh_terms: D001483:Base Sequence; D016054:DNA, Protozoan / Q000235:genetics; D004023:Dictyostelium / Q000235:genetics*; D018274:Electroporation; D004926:Escherichia coli; D055879:Gene Knock-In Techniques / Q000379:methods*; D055786:Gene Knockout Techniques / Q000379:methods*; D005822:Genetic Vectors; D059765:Homologous Recombination; D019426:Integrases / Q000235:genetics*; D008969:Molecular Sequence Data; D016254:Mutagenesis, Insertional; D010957:Plasmids / Q000235:genetics; D017422:Sequence Analysis, DNA; D014169:Transformation, Bacterial
chemical_list: D016054:DNA, Protozoan; C045073:Cre recombinase; D019426:Integrases