title: Generation of gene-targeted mice using embryonic stem cells derived from a transgenic mouse model of Alzheimer's disease.
abstract: Gene-targeting technology using mouse embryonic stem (ES) cells has become the "gold standard" for analyzing gene functions and producing disease models. Recently, genetically modified mice with multiple mutations have increasingly been produced to study the interaction between proteins and polygenic diseases. However, introduction of an additional mutation into mice already harboring several mutations by conventional natural crossbreeding is an extremely time- and labor-intensive process. Moreover, to do so in mice with a complex genetic background, several years may be required if the genetic background is to be retained. Establishing ES cells from multiple-mutant mice, or disease-model mice with a complex genetic background, would offer a possible solution. Here, we report the establishment and characterization of novel ES cell lines from a mouse model of Alzheimer's disease (3xTg-AD mouse, Oddo et al. in Neuron 39:409-421, 2003) harboring 3 mutated genes (APPswe, TauP301L, and PS1M146V) and a complex genetic background. Thirty blastocysts were cultured and 15 stable ES cell lines (male: 11; female: 4) obtained. By injecting these ES cells into diploid or tetraploid blastocysts, we generated germline-competent chimeras. Subsequently, we confirmed that F1 mice derived from these animals showed similar biochemical and behavioral characteristics to the original 3xTg-AD mice. Furthermore, we introduced a gene-targeting vector into the ES cells and successfully obtained gene-targeted ES cells, which were then used to generate knockout mice for the targeted gene. These results suggest that the present methodology is effective for introducing an additional mutation into mice already harboring multiple mutated genes and/or a complex genetic background.
authors.affiliation: Pharmaceutical Research Division, Takeda Pharmaceutical Company Limited, 2-26-1, Muraoka-Higashi, Fujisawa City, Kanagawa 251-8555, Japan. satoshi.yamamoto2@takeda.com
authors.forename: Satoshi
authors.initials: S
authors.lastname: Yamamoto
authors.forename: Yuki
authors.initials: Y
authors.lastname: Ooshima
authors.forename: Mitsugu
authors.initials: M
authors.lastname: Nakata
authors.forename: Takashi
authors.initials: T
authors.lastname: Yano
authors.forename: Kunio
authors.initials: K
authors.lastname: Matsuoka
authors.forename: Sayuri
authors.initials: S
authors.lastname: Watanabe
authors.forename: Ryouta
authors.initials: R
authors.lastname: Maeda
authors.forename: Hideki
authors.initials: H
authors.lastname: Takahashi
authors.forename: Michiyasu
authors.initials: M
authors.lastname: Takeyama
authors.forename: Yoshio
authors.initials: Y
authors.lastname: Matsumoto
authors.forename: Tadatoshi
authors.initials: T
authors.lastname: Hashimoto
mesh_terms: D000544:Alzheimer Disease / Q000235:genetics*; D016229:Amyloid beta-Peptides / Q000235:genetics; D000818:Animals; D001755:Blastocyst / Q000502:physiology; D002460:Cell Line; D004195:Disease Models, Animal; D053595:Embryonic Stem Cells / Q000502:physiology*; D018390:Gene Targeting / Q000379:methods*; D005854:Germ Cells; D006031:Glycosylation; D051379:Mice; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D008822:Mice, Transgenic; D009154:Mutation; D051381:Rats; D057891:Tetraploidy; D016875:tau Proteins / Q000235:genetics
chemical_list: D016229:Amyloid beta-Peptides; D016875:tau Proteins