| Budget Amount *help |
¥5,330,000 (Direct Cost: ¥4,100,000、Indirect Cost: ¥1,230,000)
Fiscal Year 2014: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000)
Fiscal Year 2013: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000)
Fiscal Year 2012: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000)
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| Outline of Final Research Achievements |
In a previous study, we produced mouse←rat chimera by injecting rat embryonic stem (ES) cells into mouse blastocysts. In this model, rat ES cells contributed to various organs, including spermatozoa, indicating potential utility for the production of a knockout rat. Hence, we established new rat ES cell lines to visualize rat spermatozoa using green fluorescent protein (GFP) from green body-green sperm (GBGS) doubly transgenic rats. Subsequently, we prepared a homologous recombined rat ES cell lines. When GBGS-rat ES cells were used to produce mouse←rat chimera, rat spermatozoa were easily detected with GFP in chimeric testes. Subsequently, the germline potential of rat spermatozoa in the chimera were assessed by intracytoplasmic injections. Rat pups were generated from rat spermatozoa in chimeric testes, and normal germline potential of rat spermatozoa in the chimeric testes was demonstrated. This study provides a novel method for the production of knockout rat.
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