Grant-in-Aid for Scientific Research (A)
|Allocation Type||Single-year Grants|
Laboratory animal science
|Research Institution||Kumamoto University|
SUZUKI Misao Kumamoto University, Center for Animal Resources and Development, Assintant Professor, 動物資源開発研究センター, 助教授 (60253720)
TSUJI Masayoshi Kyudo Co., Ltd., Planning Office, Laboratory Head, 企画室, 室長
TSUKUMI Kiyoshi Kumamoto University, School of Medicine, Assistant, 医学部, 助手 (60188521)
|Project Period (FY)
1996 – 1998
Completed(Fiscal Year 1998)
|Budget Amount *help
¥15,800,000 (Direct Cost : ¥15,800,000)
Fiscal Year 1998 : ¥4,500,000 (Direct Cost : ¥4,500,000)
Fiscal Year 1997 : ¥5,000,000 (Direct Cost : ¥5,000,000)
Fiscal Year 1996 : ¥6,300,000 (Direct Cost : ¥6,300,000)
|Keywords||Cre / IoxP / transgenic mice / KO mice / Embryo Freezing / BAC transgenesis / homologous recombination / ES cells / backup system in mutant mouse technology / neo耐性マウス|
In light of the great contribution of mutant technology in mice to life science, the establishment of National Center has been eagerly awaited by researchers in this field. Toward this long range goal, this study has aimed to establish its technical bases. Fortunately, Center for Animal Resources and Development is instituted in Kumamoto University and going to function fully at 2000. The fruits of this study will be transmitted to the Center.
1. Development ofgenetic technology in mice
a) The effects of insulator sequences have been intensively examined on the efficiency of transgene expressions in transgenic mice generated by DNA injection into zygotes.
b) A series of Cre transgenic mice have been generated that exhibit an unique expression spaiotemporally. These mice will serve as precious resources for conditional mutations in mice.
c) Transgenic mice that harbor large DNA cloned with BAC vector has been efficintely generated, and the modificaton of the large transgene has been success
fuly accomplished by homologous recombination in zygotes.
d) The efficiency of the isolation of homologous recombinants in ES cells has been greatly improved, and the identification of the recombinants was greatly simplified using long PCR over several kb.
e) The technology of freezing, storage in frozen, transportation in frozen and thawing of mouse zygotes, embryos and sperm have been greatly improved.
2. Technical guidance
a) Production of transgenic mice by DNA injection into zygotes
b) Construction of targeting vector
c) Culture of ES cells and isolation of homologus recombinants
d) Production of chimeric mice by injection of ES cells to embryos
e) Freezing and thawing of mouse embryos
These technologies were transmitted to. Kyudo Co. Ltd. to promote the activity in commercial base. The technologies have been also transmitted to researchers in Universuty and Research Institutes upon request.
3. Material supply
a) Transgenic mice and chimeric mice were generated for researches upon request.
b) ES cells, neo-resistant primary cells and a series of vectors have been distributed to researchers upon request. Less