| Project/Area Number |
15K01693
|
|---|
| Research Category |
Grant-in-Aid for Scientific Research (C)
|
| Allocation Type | Multi-year Fund |
|---|
| Section | 一般 |
|---|
| Research Field |
Applied health science
|
|---|
| Research Institution | Chubu University |
|---|
Principal Investigator |
AOYAMA Yuka 中部大学, 臨床検査技術教育・実習センター, 講師 (40460498)
|
|---|
| Project Period (FY) |
2015-04-01 – 2019-03-31
|
| Project Status |
Completed (Fiscal Year 2018)
|
| Budget Amount *help |
¥4,420,000 (Direct Cost: ¥3,400,000、Indirect Cost: ¥1,020,000)
Fiscal Year 2017: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000)
Fiscal Year 2016: ¥1,040,000 (Direct Cost: ¥800,000、Indirect Cost: ¥240,000)
Fiscal Year 2015: ¥2,080,000 (Direct Cost: ¥1,600,000、Indirect Cost: ¥480,000)
|
|---|
| Keywords | 先天代謝異常症 / ケトン体代謝 / β-ケトチオラーゼ / ACAT1 / スプライシング異常 / minigene splice実験 / HSD10病 / 先天性ケトン体代謝異常症 / T2欠損症 / ACAT1遺伝子 / HSD17B10遺伝子 / HSD17B10 / スプライシング / Minigene splicing 実験 |
|---|
| Outline of Final Research Achievements |
In this study, mutational analysis of an Indian T2-deficient patient revealed a homozygous c.121-13T>A mutation located at the polypyrimidine tract of the splice acceptor site of intron 2, and exon 3 skipping was identified by cDNA analysis using cycloheximide. I made the c.121-13T>A, T>C, and T>G mutant constructs followed by making a wild-type minigene construct that included an ACAT1 segment from exon 2 to 4 for a splicing experiment. The minigene splicing experiment demonstrated that exon 3 skipping was induced not only by c.121-13T>A mutation, but also by the other two substitutions. The c.121-13 position of ACAT1 gene appears to be an originally low-recognized site. In the routine diagnostic practice, in silico tools can predict the potential consequences of mutations on splicing, but their results are not so reliable. The minigene splicing experiment remains the most reliable method to unravel splicing abnormalities.
|
| Academic Significance and Societal Importance of the Research Achievements |
T2欠損症とHSD10病に関する遺伝子解析は、アジアの国々ではまだ研究が進んでいない国もあり診断が難しい状況にある。本研究では、ACAT1遺伝子のスプライスアクセプター部位のポリピリミジン領域存在するc.121-13T>Aのホモ接合性の変異がT2欠損症を引き起こすことをminigene splicing実験にて明らかにした。これら実験系の確立およびスプライシング異常を引き起こす変異の同定は、T2欠損症だけでなく他疾患においても診断の助けとなるはずである。
|
