Generation of multiple knockout mutants using the Cre-loxP system.
Dictyostelium discoideum is an exceptionally powerful system by which to study various aspects of modern cell and developmental biology. The completion of the genome sequencing project together with a high-efficiency of targeted gene disruption have enabled researchers to characterize many specific gene functions. However, as a result of many gene products with overlapping functions, there is great need to produce mutants carrying mutations in multiple genes. We have, therefore, developed a robust system and describe a protocol for the generation of multiple gene mutations in Dictyostelium by recycling the Blasticidin S selectable marker after transient expression of Cre recombinase.
Kimmel
Alan R
AR
Laboratory of Cellular and Developmental Biology, NIDDK, National Institutes of Health, Bethesda, MD, USA.
Faix
Jan
J
D000818:Animals; D030801:Animals, Genetically Modified; D004023:Dictyostelium / Q000187:drug effects / Q000235:genetics*; D017353:Gene Deletion; D018390:Gene Targeting / Q000379:methods*; D017125:Genes, Protozoan; D005822:Genetic Vectors; D019426:Integrases / Q000378:metabolism*; D009154:Mutation; D011995:Recombination, Genetic
C045073:Cre recombinase; D019426:Integrases