| Project/Area Number |
18K19579
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| Research Category |
Grant-in-Aid for Challenging Research (Exploratory)
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| Allocation Type | Multi-year Fund |
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| Review Section |
Medium-sized Section 55:Surgery of the organs maintaining homeostasis and related fields
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| Research Institution | Osaka University |
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Principal Investigator |
Miyagawa Shuji 大阪大学, 微生物病研究所, 招へい研究員 (90273648)
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| Project Period (FY) |
2018-06-29 – 2021-03-31
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| Project Status |
Completed (Fiscal Year 2020)
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| Budget Amount *help |
¥6,370,000 (Direct Cost: ¥4,900,000、Indirect Cost: ¥1,470,000)
Fiscal Year 2020: ¥2,080,000 (Direct Cost: ¥1,600,000、Indirect Cost: ¥480,000)
Fiscal Year 2019: ¥2,080,000 (Direct Cost: ¥1,600,000、Indirect Cost: ¥480,000)
Fiscal Year 2018: ¥2,210,000 (Direct Cost: ¥1,700,000、Indirect Cost: ¥510,000)
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| Keywords | PERV-C / CRISPR/Cas3 / ブタ繊維芽細胞 / PERV / ブタ線維芽細胞 / Dol-P-Man / CRISPR / ブタ内在性レトロウイルス / 再生医療 / バイオ人工臓器 / ノックアウト |
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| Outline of Final Research Achievements |
First, aiming to establish a new method of CRISPR/Cas3 in porcine cells, we set two sites in Exon9 of Gal-T (GGTA1), and tried KO using porcine fibroblasts. We introduced pX-Cascade, BPNLS-hCas3 and pBS-U6-crRNA at the same time, and confirmed a del of 294-754 bp in Sequence. Furthermore, Cas3-crRNA was produced at two sites for HD CMAH.
Next, Cas3-crRNA for Dol-P-mannosidase was also created. Moreover, as to KO PERV-C, Cas3-crRNA was prepared on the TM side of enb in the A/C chimera, and its effect on porcine fibroblasts was examined.
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| Academic Significance and Societal Importance of the Research Achievements |
特許が障害となっているCRISPR/Cas9法ではなく、日本発の新法であるCRISPR/Cas3法がブタの細胞でも有効であることが証明された。また、ブタPERVのKOに関しても、この方法が利用できることが期待される。
これにより、PERV感染の可能性の少ないブタが作出され、再生医療やバイオ人工臓器に使われれば、現実の医療を変える効果が期待できる。
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