title: In vivo evaluation of ΦC31 recombinase activity in transgenic mice.
abstract: Genome engineering strategies employing site-specific recombinases (SSRs) such as Cre, Flp and PhiC31, have become powerful tools to analyze gene function and manipulate neural network in vertebrates. In the present study, we evaluated the ability of PhiC31 phage integrase to induce genomic recombination in transgenic mice. PhiC31 is the integrase encoded by the Streptomyces bacteriophage that promotes recombination between heterotypic attP and attB sites. We generated transgenic mice that express codon-optimized PhiC31 (PhiC31o) in neural stem/progenitor cells or tyrosine hydroxylase (TH) expressing catecholaminergic neurons. PhiC31 was functional in these cells and capable of excising a transcriptional stop cassette flanked by PhiC31-specific attP/B recognition sites. PhiC31-ER(T2), a fusion protein of PhiC31o (without the nuclear localization signal) and the mutated ligand-binding domain of the human estrogen receptor, was able to induce recombination in neural stem/progenitor cells in a tamoxifen-dependent manner, but the recombination rate was less efficient than for PhiC31. Thus, PhiC31 integrase is functional in transgenic mice and is suitable for mosaic recombination in restricted cell populations.
authors.affiliation: Institute for Virus Research, Kyoto University, Kyoto 606-8507, Japan. iimayosh@virus.kyoto-u.ac.jp
authors.forename: Itaru
authors.initials: I
authors.lastname: Imayoshi
authors.forename: Kyoko
authors.initials: K
authors.lastname: Hirano
authors.forename: Satsuki
authors.initials: S
authors.lastname: Kitano
authors.forename: Hitoshi
authors.initials: H
authors.lastname: Miyachi
authors.forename: Ryoichiro
authors.initials: R
authors.lastname: Kageyama
mesh_terms: D000818:Animals; D001435:Bacteriophages / Q000201:enzymology* / Q000235:genetics*; D004789:Enzyme Activation / Q000502:physiology; D005260:Female; D005818:Genetic Engineering / Q000379:methods / Q000592:standards*; D008297:Male; D051379:Mice; D008822:Mice, Transgenic; D058953:Neural Stem Cells / Q000201:enzymology; D045522:Recombinases / Q000235:genetics* / Q000378:metabolism*; D011995:Recombination, Genetic
chemical_list: D045522:Recombinases