2013 Fiscal Year Final Research Report
Improved technology of mouse somatic cell nuclear transfer that greatly increases cloning efficiency
| Project/Area Number |
23580390
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Multi-year Fund |
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| Section | 一般 |
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| Research Field |
Applied animal science
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| Research Institution | Kyoto University |
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Principal Investigator |
YAMADA Masayasu 京都大学, (連合)農学研究科(研究院), 准教授 (10243073)
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| Project Period (FY) |
2011 – 2013
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| Keywords | 体細胞核移植 / クローンマウス / ヒストン脱アセチル化酵素阻害剤 / 脱イオン化BSA / ビタミンC / エピジェネティックス / X染色体不活化異常 |
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| Research Abstract |
Production rate of cloned mice still is very low, which is thought to be attributed to abnormal epigenetic modifications and X-chromosomal inactivation (XCI) status in somatic cell nuclear transfer (SCNT) embryos. Here, we developed novel mouse SCNT technologies as follows; 1) treatment with valproi acid for 24 h from the 4-cell stage of SCNT embryos, which improved expression of Oct4 and trimethylated histone H3 lysine 27, a marker of the XCI status in SCNT blastocysts, 2) intracytoplasmic injection of deionized BSA after SCNT, which promoted expression of acetylated histone H3 lysine 9 and acetylated histone H4 lysine 12 at the pronuclear stage and production of cloned offspring, and 3) treatments with tricostatine A for 8 h from the beginning of activation of SCNT oocytes followed by vitamin C for 7 h, which promoted the oxidation of 5-methylcytocine to 5-hydroxymethylcytosine in pronuclear DNA of SCNT embryos and led to greatly increased production of cloned offspring.
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[Journal Article] Identification and characterization of an oocyte factor required for development of porcine nuclear transfer embryos2011
Author(s)
Miyamoto K, Nagai K, Kitamura N, Nishikawa T, Ikegami H, Binh NT, Tsukamoto S, Matsumoto M, Tsukiyama T, Minami N, Yamada M, Ariga H, Miyake M, Kawarasaki T, Matsumoto K, Imai H.
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Journal Title
Proc. Natl. Acad. Sci. USA
Volume: 108(17)
Pages: 7040-7045
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