Development of transgenic animals using a human artificial chromosome (HAC) vector

• Project/Area Number: 22580320
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Applied animal science
• Research Institution: Yamaguchi University
• Principal Investigator: OTOI Takeshige 山口大学, 共同獣医学部, 教授 (30311814)
• Project Period (FY): 2010 – 2012
• Project Status: Completed (Fiscal Year 2012)
• Budget Amount: ¥4,550,000 (Direct Cost: ¥3,500,000、Indirect Cost: ¥1,050,000); Fiscal Year 2012: ¥780,000 (Direct Cost: ¥600,000、Indirect Cost: ¥180,000); Fiscal Year 2011: ¥1,040,000 (Direct Cost: ¥800,000、Indirect Cost: ¥240,000); Fiscal Year 2010: ¥2,730,000 (Direct Cost: ¥2,100,000、Indirect Cost: ¥630,000)
• Keywords: 遺伝子改変動物 / 人工染色体 / クローン / 蛍光タンパク質 / ネコ / 胚発育 / ベクター

Research Abstract

We established the two cell lines of cat fibroblasts with a human artificial chromosome (HAC) vector containing the green fluorescent protein (GFP). By inserting the transgenic cell into an enucleated cat egg, the cloned embryos were produced. The ability for cloned embryos to develop to blastocysts was higher in the short-time fusion group (30 μsec) than in the long-time fusion group (60 μsec). The embryos were transferred to 13 recipients. Only one recipient became pregnant and delivered a live kitten (74g).

Research Products

• [Journal Article] Histone deacetylase inhibitor improves the development and acetylation levels of cat cow interspecies cloned embryos (2013)
  • Author(s): Wittayarat, M., Sato, Y., Do, T.K.L., Morita, Y., Chatdarong, K., Techakumphu, M., Taniguchi, M. and Otoi, T.
  • Journal Title: Cellular Reprogramming
  • Peer Reviewed
• [Journal Article] Cell cycle analysis and interspecies nuclear transfer of cat cells treated with chemical inhibitors (2013)
  • Author(s): Wittayarat, M., Fujiwara, A., Chatdarong, K., Techakumphu, M., Sato, Y., Tanihara, F., Morita, Y., Taniguchi, M. and Otoi, T
  • Journal Title: Acta Veterinaria Hungarica
  • Peer Reviewed
• [Journal Article] The effect of relaxin supplementation of in vitro maturation medium on the development of cat oocytes obtained from ovaries stored at 4 °C (2013)
  • Author(s): Luu, V.V., Hanatate, K., Tanihara, F., Sato, Y., Do, T.K.L., Taniguchi, M., and Otoi, T.
  • Journal Title: Reproductive Biology
  • Peer Reviewed
• [Journal Article] Comparison of activation ability between feline and bovine oocytes (2013)
  • Author(s): Tanihara, F., Kaedei, Y., Namula, Z., Luu, V.V., Sato, Y., Wittayarat, M., Taniguchi, M. and Otoi, T
  • Journal Title: Acta Veterinaria Hungarica
  • Peer Reviewed
• [Journal Article] Cell cycle synchronization of skin fibroblast cells in four species of family Felidae (2013)
  • Author(s): Wittayarat, M., Thongphakdee, A., Saikhun, K., Chatdarong, K., Otoi, T.and Techakumphu, M
  • Journal Title: Reproduction in Domestic Animals Volume: 48 Pages: 305-310
  • Peer Reviewed
• [Journal Article] Cell cycle synchronization of skin fibroblast cells in four species of family Felidae. (2013)
  • Author(s): Wittayarat, M.
  • Journal Title: Reproduction in Domestic Animals Volume: 48 Issue: 2 Pages: 305-310
  • DOI: 10.1111/j.1439-0531.2012.02149.x
  • Peer Reviewed
• [Journal Article] Development and subsequent cryotolerance of domestic cat embryos cultured in serum-free and serum-containing media (2011)
  • Author(s): Murakami, M., Dong, Y.J., Suzuki, T., Taniguchi, M., Kaedei, Y., Sato, Y., Tanihara, F. and Otoi, T
  • Journal Title: Cryobiology Volume: 63 Pages: 170-174
  • Peer Reviewed
• [Journal Article] Antiviral restriction factor transgenesis in the domestic cat. (2011)
  • Author(s): Wongsrikeao, P., Saenz, D., Rinkoski, T., Otoi, T. and Poeschla, E
  • Journal Title: Nature Methods Volume: 8 Pages: 853-859
  • Peer Reviewed
• [Journal Article] Antiviral restriction factor transgenesis in the domestic cat (2011)
  • Author(s): Wongsrikeao, P., et al
  • Journal Title: Nature Methods Volume: 8 Issue: 10 Pages: 853-859
  • DOI: 10.1038/nmeth.1703
  • Peer Reviewed
• [Journal Article] In vitro development of cat interspecies nuclear transfer using pig's and cow's cytoplasm. (2010)
  • Author(s): Kaedei, Y.
  • Journal Title: B.Vet.I.Pulawy Volume: 54 Pages: 405-408
  • Peer Reviewed
• [Presentation] TEffect of trichostatin A on in vitro embryo development of interspecies nuclear transfer embryos reconstructed from cat donor nuclei and bovine cytoplasm. 39th Annual Conference of the IETS (2013)
  • Author(s): Wittayarat, M., Namula, Z., Luu, V. V., Do, L. T. K., Sato, Y., Taniguchi, M. and Otoi
  • Organizer: 39th Annual Conference of the IETS
  • Place of Presentation: Hannover, Germany
  • Year and Date: 2013-01-20
• [Presentation] Effects of ovary storage time on the quality and meiotic competence of cat oocytes (2012)
  • Author(s): Luu, V.V., et al
  • Organizer: 38回国際胚移植学会
  • Place of Presentation: Phoenix AZ USA
  • Year and Date: 2012-01-09
• [Presentation] Effects of ovary storage time on the quality and meiotic competence of cat oocytes (2012)
  • Author(s): Luu, V. V., Namula, Z., Kaedei, Y., Tanihara, F. and Otoi, T
  • Organizer: 38th Annual Conference of the IETS
  • Place of Presentation: Phoenix, AZ, USA
  • Year and Date: 2012-01-08
• [Presentation] ネコ卵母細胞の体外成熟・体外発育に及ぼすリラキシンの添加効果 (2010)
  • Author(s): 花立敬祐
  • Organizer: 150回日本獣医学会
  • Place of Presentation: 帯広市、帯広畜産大学
  • Year and Date: 2010-09-16
• [Presentation] 異なる機序の細胞周期同調剤で処理したネコ体細胞と除核ウシ卵母細胞との融合後の胚発育 (2010)
  • Author(s): 藤原彬
  • Organizer: 150回日本獣医学会
  • Place of Presentation: 帯広市、帯広畜産大学
  • Year and Date: 2010-09-16
• [Presentation] Effect of trichostatin A on in vitro embryo development of interspecies nuclear transfer embryos reconstructed from cat donor nuclei and bovine cytoplasm.
  • Author(s): Wittayarat, M.
  • Organizer: 39th Annual Conference of the IETS
  • Place of Presentation: Hannover, Germany