| Project/Area Number |
25660011
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| Research Category |
Grant-in-Aid for Challenging Exploratory Research
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| Allocation Type | Multi-year Fund |
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| Research Field |
Science in genetics and breeding
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| Research Institution | National Institute of Advanced Industrial Science and Technology |
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Principal Investigator |
MITSUDA Nobutaka 独立行政法人産業技術総合研究所, 生物プロセス研究部門, 主任研究員 (80450667)
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| Co-Investigator(Kenkyū-buntansha) |
KATO Yoshio 国立研究開発法人産業技術総合研究所, 生命工学領域・バイオメディカル研究部門, 主任研究員 (20415657)
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| Project Period (FY) |
2013-04-01 – 2015-03-31
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| Project Status |
Completed (Fiscal Year 2014)
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| Budget Amount *help |
¥3,900,000 (Direct Cost: ¥3,000,000、Indirect Cost: ¥900,000)
Fiscal Year 2014: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000)
Fiscal Year 2013: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000)
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| Keywords | CRES-T / 転写抑制 / 転写因子 / CRISPR / 植物 / 発現抑制 / 遺伝子発現 |
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| Outline of Final Research Achievements |
To determine which region is most effective to suppress expression of target gene by chimeric repressor, repeated GAL4 binding sequence was inserted into LUC gene. However, it was found that LUC activity becomes much reduced even without any effector in that case. We therefore decided to employ CRISPR-Cas9 system for our purpose. We prepared vector that expresses gRNA and mutated CAS9 (D10A H840A) fused with SRDX which doesn’t digest DNA. As a test experiment, we designed several gRNAs for the reporter construct 35S:GAL4UASx5:TATA:LUC, however, we didn’t see any repression of the reporter when we expressed these gRNAs and mutated CAS9 fused with SRDX together at least in the level of transient assay.
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