| Project/Area Number |
20K06248
|
|---|
| Research Category |
Grant-in-Aid for Scientific Research (C)
|
| Allocation Type | Multi-year Fund |
|---|
| Section | 一般 |
|---|
| Review Section |
Basic Section 40040:Aquatic life science-related
|
|---|
| Research Institution | Fisheries Research and Education Agency |
|---|
Principal Investigator |
Kawato Yasuhiko 国立研究開発法人水産研究・教育機構, 水産技術研究所(南勢), 主任研究員 (90634220)
|
|---|
| Project Period (FY) |
2020-04-01 – 2023-03-31
|
| Project Status |
Completed (Fiscal Year 2022)
|
| Budget Amount *help |
¥4,290,000 (Direct Cost: ¥3,300,000、Indirect Cost: ¥990,000)
Fiscal Year 2022: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000)
Fiscal Year 2021: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000)
Fiscal Year 2020: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000)
|
|---|
| Keywords | ゲノム編集 / ワクチン / 複製阻害 / マダイイリドウイルス / 培養細胞 / ウイルス / CRISPR-Cas9 / ウイルス複製阻害因子 / 水産学 |
|---|
| Outline of Research at the Start |
本研究では、ゲノム編集によるマダイイリドウイルスの効率的なワクチン製造が可能な培養細胞株の樹立を目指す。網羅的な遺伝子発現解析によりウイルス複製阻害因子を推定し、CRISPR/Cas9によるゲノム編集を利用して当該因子をノックアウトする。薬剤マーカーによるスクリーニングでノックアウト細胞株を安定化させ、マダイイリドウイルスの増殖性を確認する。
|
| Outline of Final Research Achievements |
In this study, we attempted to establish a cell line in which viral replication inhibitory factors were knocked out using genome editing technology for the purpose of efficient production of inactivated viral vaccines. Transcriptome analysis using a cell line derived from spotted knifejaw Oplegnathus punctatus which is used to propagate red sea bream iridovirus suggested that a gene cluster involved in an immune response may be involved in the inhibition of virus replication. On the basis of the information, the genome editing using the CRISPR-Cas9 system was conducted targeting two genes. Although we were able to maintain cells in which the gene was knocked out for approximately one month, we were not able to evaluate the viral susceptibility of the knockout cells because the cell growth was very slow.
|
| Academic Significance and Societal Importance of the Research Achievements |
培養細胞におけるマダイイリドウイルスの複製阻害要因が推定され、当該遺伝子をノックアウトする系も構築されつつある。ノックアウト細胞のスクリーニングと培養条件が整理できれば、ゲノム編集細胞株を用いた効率的なワクチン製造の足掛かりとなる可能性がある。
|
