| Project/Area Number |
15K11090
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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 |
Pathobiological dentistry/Dental radiology
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| Research Institution | Kanagawa Dental College |
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Principal Investigator |
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| Co-Investigator(Renkei-kenkyūsha) |
HATA Ryu-Ichiro 神奈川歯科大学, 歯学部, 特任教授 (10014276)
KATOH Iyoko 山梨大学, 総合研究部, 准教授 (20333297)
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| Project Period (FY) |
2015-04-01 – 2018-03-31
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| Project Status |
Completed (Fiscal Year 2017)
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| Budget Amount *help |
¥4,680,000 (Direct Cost: ¥3,600,000、Indirect Cost: ¥1,080,000)
Fiscal Year 2017: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000)
Fiscal Year 2016: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000)
Fiscal Year 2015: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000)
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| Keywords | p63 / CRISPR-Cas9 / ゲノム編集 |
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| Outline of Final Research Achievements |
P63(TP63), one of the tumor suppressor p53 gene family, is highly expressed in non-invasive squamous carcinomas of head-and-neck. To investigate the role of p63, we attempted genome editing with CRISPR-Cas9. P63 has TA and ΔN type transcription start sites, of which we focused on the first exon of TA-p63. We selected drug-resistant cells after transfection of carcinoma cells with a guide vector containing the gRNA and Cas9 together with a donor vector containing a drug-resistance gene between the 5’ and 3’ flanking regions of the target site. This procedure allowed recombination at the expected site, but only at a single allele. To achieve the biallelic knockout, we needed to perform step-wise transfection-and-selection cycles using two donor vectors with different drug resistance genes.
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