Direct Production of Gene-targeted Mice from ES Cells by Nuclear Transfer and Gene Transmission to their Progeny

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概要

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• In order to evaluate the usefulness of a cloning technique to produce gene-manipulated mice for the field of laboratory animal science, we produced mice cloned from gene-targeted embryonic stem (ES) cells and examined the vertical transmission of a targeted gene to their progeny. Of 1257 eggs constructed by nuclear transfer using M-phase ES donor cells targeted with an oviduct-specific glycoprotein (OGP) gene, 990 formed a pseudo-pronucleus and a polar body after activation. Of 504 cloned embryos transferred into recipients, 20 live cloned pups (2%) were recovered by Caesarean section at 19.5 days of gestation. Fourteen of these cloned mice were studied. Genotyping of the OGP locus and 20 microsatellite loci showed that they were genetically identical to the OGP gene-targeted TT2 cells. Eight cloned pups grew into adults, of which 7 were male and 1 was female (missing the Y chromosome). Mating experiments using the cloned mice were carried out. Of 89 F1 mice produced from the mating of cloned and C57BL/6J mice, 50 had the targeted OGP gene heterozygously. Thirty-seven F2 mice from 4 pairs of the OGP-/+ mice were composed of 9 OGP-/-, 18 OGP-/+, and 10 OGP+/+. Moreover, 26 offspring of one pair of the cloned mice were composed of 10 OGP-/-, 12 OGP-/+, and 4 OGP+/+. These offspring were fertile and transmitted the mutant OGP gene to the next generation. Comparison of these results with those of germline chimeric mice indicates that gene-targeted mice can be produced at least one generation earlier by nuclear transfer than by the conventional methods. In addition, the targeted OGP gene was constantly transmitted to the progeny of the gene-targeted mice. Cloning techniques are potentially a more efficient way to generate gene-manipulated mice for laboratory animal science, although such techniques include many unresolved problems, such as low production efficiency, and selection of a cell source for gene manipulation among others.

• 社団法人　日本実験動物学会の論文
• 2002-07-01

著者

• ARAKI Yoshihiko

  Department of Immunology & Parasitology, Yamagata University School of Medicine

• KONO Tomohiro

  Department of BioScience, Tokyo University of Agriculture

• ITO Mamoru

  Central Institute for Experimental Animals

• Hioki Kyoji

  Central Inst. For Experimental Animals

• Hioki K

  Central Inst. Experimental Animals Kanagawa

• Hioki K

  Central Inst. Of Experimental Animals Kawasaki Jpn

• Shinkai Yoichi

  Institute For Virus Research Kyoto University

• SHIMOZAWA Nobuhiro

  Central Institute for Experimental Animals

• ONO Yukiko

  Department of Bioscience, Tokyo University of Agriculture

• MUGURUMA Kaori

  Central Institute for Experimental Animals

• Araki Yoshihiko

  Department Of Cardiology Osaka Prefecture Medical Center For Respiratory And Allergic Disease

• Araki Yoshihiko

  Department Of Immunology & Parasitology Yamagata University School Of Medicine

• Kono T

  Tokyo Univ. Agriculture Tokyo Jpn

• Kono Tomohiro

  Department Of Animal Science Tokyo University Of Agriculture

• Shinkai Y

  Graduate School Of Natural Science And Technology Okayama University

• Muguruma K

  Central Inst. Experimental Animals Kanagawa

• Ito M

  Central Inst. For Experimental Animals

• Shimozawa N

  Central Inst. Experimental Animals Kanagawa

• Kono T

  Nodai Research Institute Tokyo University Of Agriculture

• Ito Mamoru

  Central Inst. For Experimental Animals Kawasaki Jpn

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