| Project/Area Number |
26870845
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| Research Category |
Grant-in-Aid for Young Scientists (B)
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| Allocation Type | Multi-year Fund |
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| Research Field |
Genetics/Chromosome dynamics
Developmental biology
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| Research Institution | University of Yamanashi |
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Principal Investigator |
OTA Satoshi 山梨大学, 総合研究部, 医学研究員 (60553310)
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| Project Period (FY) |
2014-04-01 – 2016-03-31
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| Project Status |
Completed (Fiscal Year 2015)
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| Budget Amount *help |
¥4,030,000 (Direct Cost: ¥3,100,000、Indirect Cost: ¥930,000)
Fiscal Year 2015: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000)
Fiscal Year 2014: ¥2,210,000 (Direct Cost: ¥1,700,000、Indirect Cost: ¥510,000)
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| Keywords | CRISPR/Cas9 / ゼブラフィッシュ / ノックイン / トランスジェニック / 遺伝子破壊 / ノックアウト / Gal4-UAS / GFP / pax2a / ゲノム編集 / コンディショナルノックアウト |
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| Outline of Final Research Achievements |
In this study, we generated transgenic zebrafish that expresses nuclease Cas9 under control of the UAS promoter. However, guide RNA injection in transgenic embryos that express GFP did not induce insertion or deletion mutations at the target locus.
We also developed a new method for analyzing gene function and expression using CRISPR/Cas9. We inserted the donor vector containing a heat-shock promoter and GFP into the 5'-untranslated region of the pax2a locus. These embryos expressed GFP at the region where endogenous pax2a expresses. We found that homozygous transgenic embryos presented the pax2a mutant phenotype. Thus, targeted insertion of the reporter gene at the endogenous gene is useful for analyzing gene expression and function.
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