title: Efficient mapping of transgene integration sites and local structural changes in Cre transgenic mice using targeted locus amplification.
abstract: Cre/LoxP technology is widely used in the field of mouse genetics for spatial and/or temporal regulation of gene function. For Cre lines generated via pronuclear microinjection of a Cre transgene construct, the integration site is random and in most cases not known. Integration of a transgene can disrupt an endogenous gene, potentially interfering with interpretation of the phenotype. In addition, knowledge of where the transgene is integrated is important for planning of crosses between animals carrying a conditional allele and a given Cre allele in case the alleles are on the same chromosome. We have used targeted locus amplification (TLA) to efficiently map the transgene location in seven previously published Cre and CreERT2 transgenic lines. In all lines, transgene insertion was associated with structural changes of variable complexity, illustrating the importance of testing for rearrangements around the integration site. In all seven lines the exact integration site and breakpoint sequences were identified. Our methods, data and genotyping assays can be used as a resource for the mouse community and our results illustrate the power of the TLA method to not only efficiently map the integration site of any transgene, but also provide additional information regarding the transgene integration events.
authors.affiliation: Department of Transgenic Technology, Genentech, Inc., 1 DNA Way, South San Francisco, CA 94080, USA.
authors.forename: Carol
authors.initials: C
authors.lastname: Cain-Hom
authors.affiliation: Cergentis BV, Yalelaan 62, 3584 CM Utrecht, the Netherlands.
authors.forename: Erik
authors.initials: E
authors.lastname: Splinter
authors.affiliation: Cergentis BV, Yalelaan 62, 3584 CM Utrecht, the Netherlands.
authors.forename: Max
authors.initials: M
authors.lastname: van Min
authors.affiliation: Cergentis BV, Yalelaan 62, 3584 CM Utrecht, the Netherlands.
authors.forename: Marieke
authors.initials: M
authors.lastname: Simonis
authors.affiliation: Cergentis BV, Yalelaan 62, 3584 CM Utrecht, the Netherlands.
authors.forename: Monique
authors.initials: M
authors.lastname: van de Heijning
authors.affiliation: Department of Transgenic Technology, Genentech, Inc., 1 DNA Way, South San Francisco, CA 94080, USA.
authors.forename: Maria
authors.initials: M
authors.lastname: Martinez
authors.affiliation: Department of Transgenic Technology, Genentech, Inc., 1 DNA Way, South San Francisco, CA 94080, USA.
authors.forename: Vida
authors.initials: V
authors.lastname: Asghari
authors.affiliation: Department of Transgenic Technology, Genentech, Inc., 1 DNA Way, South San Francisco, CA 94080, USA.
authors.forename: J Colin
authors.initials: JC
authors.lastname: Cox
authors.affiliation: Department of Molecular Biology, Genentech, Inc., 1 DNA Way, South San Francisco, CA 94080, USA.
authors.forename: Søren
authors.initials: S
authors.lastname: Warming
mesh_terms: D000818:Animals; D002874:Chromosome Mapping / Q000379:methods*; D018628:Gene Dosage; D015870:Gene Expression; D015723:Gene Library; D056426:Genetic Loci; D016678:Genome; D059014:High-Throughput Nucleotide Sequencing; D019426:Integrases / Q000235:genetics* / Q000378:metabolism; D051379:Mice; D008810:Mice, Inbred C57BL; D008822:Mice, Transgenic; D016254:Mutagenesis, Insertional; D021141:Nucleic Acid Amplification Techniques; D013154:Spleen / Q000166:cytology / Q000378:metabolism; D019076:Transgenes
chemical_list: C045073:Cre recombinase; D019426:Integrases