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Establishment and characterization of CAG/EGFP transgenic rabbit line

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Abstract

Cell marking is a very important procedure for identifying donor cells after cell and/or organ transplantation in vivo. Transgenic animals expressing marker proteins such as enhanced green fluorescent protein (EGFP) in their tissues are a powerful tool for research in fields of tissue engineering and regenerative medicine. The purpose of this study was to establish transgenic rabbit lines that ubiquitously express EGFP under the control of the cytomegalovirus immediate early enhancer/beta-actin promoter (CAG) to provide a fluorescent transgenic animal as a bioresource. We microinjected the EGFP expression vector into 945 rabbit eggs and 4 independent transgenic candidate pups were obtained. Two of them died before sexual maturation and one was infertile. One transgenic male candidate founder rabbit was obtained and could be bred by artificial insemination. The rabbit transmitted the transgene in a Mendelian manner. Using fluorescence in situ hybridization analysis, we detected the transgene at 7q11 on chromosome 7 as a large centromeric region in two F1 offspring (one female and one male). Eventually, one transgenic line was established. Ubiquitous EGFP florescence was confirmed in all examined organs. There were no gender-related differences in fluorescence. The established CAG/EGFP transgenic rabbit will be an important bioresource and a useful tool for various studies in tissue engineering and regenerative medicine.

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Acknowledgments

We greatly acknowledge Mr Kazuo Hirasawa, president of Kitayama Labes Co. Ltd, for providing the experimental facilities and animals. We would like to thank Mr Kensaku Kitada, Mr Masao, Hirao Kitayama Labes Co. Ltd, for the skillful technical assistance. We would also like to thank Dr Kazuo Gotoh, The Central Institute for Experimental Animals, for his help in FISH analysis of the transgene, and Mr Akira Shiota, the YS institute Inc, for help in genotype analysis of transgenic rabbit.

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Author notes

  1. Shu Hashimoto

    Present address: IVF Namba Clinic, 1-17-28 Minamihorie, Nishi-ku, Osaka, Japan

  2. Yoji Hakamata

    Present address: Department of Basic Science, School of Veterinary Nursing and Technology, Nippon Veterinary and Life Science University, 1-7-1, Kyonan-cho, Musashino-shi, Tokyo, Japan

Authors and Affiliations

  1. The YS Institute Inc., 1198-4 Iwazo, Utsunomiya, Tochigi, Japan

    Ri-ichi Takahashi, Takashi Kuramochi, Kazuki Aoyagi, Shu Hashimoto & Masatsugu Ueda

  2. Center for Experimental Animal Science, Nagoya City University Graduate School of Medical Sciences, 1 Kawasumi, Mizuho-cho, Mizuho-ku, Nagoya, Japan

    Ichiro Miyoshi

  3. Institute for Animal experimentation, Tohoku University Graduate School of Medicine, 2-1 Seriyo-cho, Aoba-ku, Sendai, Japan

    Noriyuki Kasai

  4. Division of Organ Replacement Research, Center for Molecular Medicine, Jichi Medical School, 3311-1 Yakushiji, Shimotsuke, Tochigi, Japan

    Yoji Hakamata & Eiji Kobayashi

Authors
  1. Ri-ichi Takahashi
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  2. Takashi Kuramochi
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  3. Kazuki Aoyagi
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  4. Shu Hashimoto
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  8. Eiji Kobayashi
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  9. Masatsugu Ueda
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Correspondence to Ri-ichi Takahashi.

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Takahashi, Ri., Kuramochi, T., Aoyagi, K. et al. Establishment and characterization of CAG/EGFP transgenic rabbit line. Transgenic Res 16, 115–120 (2007). https://doi.org/10.1007/s11248-006-9043-1

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  • DOI: https://doi.org/10.1007/s11248-006-9043-1

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