Control of porcine endogenous retrovirus for regenerative medicine using pigs as a scaffold and development of bio-organs

2020 Fiscal Year Final Research Report

• Project/Area Number: 18K19579
• Research Category: Grant-in-Aid for Challenging Research (Exploratory)
• Allocation Type: Multi-year Fund
• Review Section: Medium-sized Section 55:Surgery of the organs maintaining homeostasis and related fields
• Research Institution: Osaka University
• Principal Investigator: Miyagawa Shuji 大阪大学, 微生物病研究所, 招へい研究員 (90273648)
• Project Period (FY): 2018-06-29 – 2021-03-31
• Keywords: PERV-C / CRISPR/Cas3 / ブタ繊維芽細胞

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.

Free Research Field

移植免疫

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。