title: Binary recombinase systems for high-resolution conditional mutagenesis.
abstract: Conditional mutagenesis using Cre recombinase expressed from tissue specific promoters facilitates analyses of gene function and cell lineage tracing. Here, we describe two novel dual-promoter-driven conditional mutagenesis systems designed for greater accuracy and optimal efficiency of recombination. Co-Driver employs a recombinase cascade of Dre and Dre-respondent Cre, which processes loxP-flanked alleles only when both recombinases are expressed in a predetermined temporal sequence. This unique property makes Co-Driver ideal for sequential lineage tracing studies aimed at unraveling the relationships between cellular precursors and mature cell types. Co-InCre was designed for highly efficient intersectional conditional transgenesis. It relies on highly active trans-splicing inteins and promoters with simultaneous transcriptional activity to reconstitute Cre recombinase from two inactive precursor fragments. By generating native Cre, Co-InCre attains recombination rates that exceed all other binary SSR systems evaluated in this study. Both Co-Driver and Co-InCre significantly extend the utility of existing Cre-responsive alleles.
authors.affiliation: Institute of Laboratory Animal Science, University of Zurich, Sternwartstrasse 6, CH-8091 Zurich, Switzerland, Institute of Neuropathology, University Hospital of Zurich, Schmelzbergstrasse 12, CH-8091 Zurich, Switzerland, Institute of Pharmacology and Toxicology, University of Zurich, Winterthurerstrasse 190, CH-8057 Zurich, Switzerland, National Centre for Biotechnology (CNB-CSIC), Darwin 3, 28049 Madrid, Spain, CIBERER-ISCIII, Darwin 3, 28049 Madrid, Spain, Program of Cardiovascular Development, Department of Cardiovascular Development and Repair, Centro Nacional de Investigaciones Cardiovasculares Carlos III, calle Melchor Fernández Almagro 3, 28029 Madrid, Spain and Institute of Pharmaceutical Sciences, Swiss Federal Institute of Technology (ETH) Zurich, Winterthurerstrasse 190, CH-8057 Zurich, Switzerland.
authors.forename: Mario
authors.initials: M
authors.lastname: Hermann
authors.forename: Patrick
authors.initials: P
authors.lastname: Stillhard
authors.forename: Hendrik
authors.initials: H
authors.lastname: Wildner
authors.forename: Davide
authors.initials: D
authors.lastname: Seruggia
authors.forename: Viktor
authors.initials: V
authors.lastname: Kapp
authors.forename: Héctor
authors.initials: H
authors.lastname: Sánchez-Iranzo
authors.forename: Nadia
authors.initials: N
authors.lastname: Mercader
authors.forename: Lluís
authors.initials: L
authors.lastname: Montoliu
authors.forename: Hanns Ulrich
authors.initials: HU
authors.lastname: Zeilhofer
authors.forename: Pawel
authors.initials: P
authors.lastname: Pelczar
mesh_terms: D000818:Animals; D001921:Brain / Q000378:metabolism; D002460:Cell Line; D017930:Genes, Reporter; D057809:HEK293 Cells; D006801:Humans; D019426:Integrases / Q000378:metabolism*; D051379:Mice; D016296:Mutagenesis; D019579:Neocortex / Q000378:metabolism; D045522:Recombinases / Q000378:metabolism*; D011995:Recombination, Genetic
chemical_list: D045522:Recombinases; C045073:Cre recombinase; D019426:Integrases