title: The Cre/loxP system in Giardia lamblia: genetic manipulations in a binucleate tetraploid protozoan.
abstract: The bacteriophage-derived Cre/loxP system is a valuable tool that has revolutionised genetic and cell biological research in many organisms. We implemented this system in the intestinal parasite Giardia lamblia, an evolutionarily diverged protozoan whose binucleate and tetraploid genome organisation severely limits the application of reverse genetic approaches. We show that Cre-recombinase is functionally expressed in G. lamblia and demonstrate "recycling" of selectable markers. Providing the means for more complex and versatile genetic modifications, this technique massively increases the scope of functional investigations in G. lamblia and other protozoa with similar limitations with respect to genetic manipulation.
authors.affiliation: Laboratory of Molecular Parasitology, Institute of Parasitology-University of Zurich, Winterthurerstrasse 266a, CH-8057 Zürich, Switzerland.
authors.forename: Petra B
authors.initials: PB
authors.lastname: Wampfler
authors.affiliation: Laboratory of Molecular Parasitology, Institute of Parasitology-University of Zurich, Winterthurerstrasse 266a, CH-8057 Zürich, Switzerland.
authors.forename: Carmen
authors.initials: C
authors.lastname: Faso
authors.affiliation: Laboratory of Molecular Parasitology, Institute of Parasitology-University of Zurich, Winterthurerstrasse 266a, CH-8057 Zürich, Switzerland. Electronic address: adrian.hehl@uzh.ch.
authors.forename: Adrian B
authors.initials: AB
authors.lastname: Hehl
mesh_terms: D001435:Bacteriophages / Q000201:enzymology / Q000235:genetics; D015870:Gene Expression; D005827:Genetics, Microbial / Q000379:methods*; D016829:Giardia lamblia / Q000235:genetics*; D019426:Integrases / Q000235:genetics / Q000378:metabolism; D008967:Molecular Biology / Q000379:methods*; D011995:Recombination, Genetic; D057891:Tetraploidy
chemical_list: C045073:Cre recombinase; D019426:Integrases
keywords: Cre/loxP; Genetic manipulation; Giardia; Resistance marker recycling; Selectable marker; Tetraploid