2015 Fiscal Year Final Research Report
Development of mutagenesis to the mitochondrial DNA using artificial restriction enzyme and application to definitive diagnosis.
| Project/Area Number |
25670261
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| Research Category |
Grant-in-Aid for Challenging Exploratory Research
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| Allocation Type | Multi-year Fund |
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| Research Field |
Applied pharmacology
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| Research Institution | Saitama Medical University |
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Principal Investigator |
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| Project Period (FY) |
2013-04-01 – 2016-03-31
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| Keywords | ミトコンドリアDNA / 遺伝子診断・治療 / 人工制限酵素 / 遺伝子工学 |
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| Outline of Final Research Achievements |
We performed whole mitochondrial DNA (mtDNA) sequence analysis in patients with mitochondrial respiratory chain disorder, and identified several plausible causative mutations. Here, we tried to produce mutant cell lines harboring the variants of unknown significance (VUSs) from normal cells using artificial nucleases in the aim for establishing a new method for definitive diagnosis for VUSs.
We applied TALENs (Transcription activator-like effector nucleases) system, and engineered so that they localize to mitochondria and recognize the specific target sequence on mtDNA. Actually, the engineered mitochondria-targeted TALENs correctly recognized one-base -difference in the specific target sequence, and cut the proper site.
Although we’ve not achieved producing mutant cell lines from normal cells, our TALEN system cut the specific mtDNA site. This system can control the heteroplasmy rate inside the mitochondria, and will pave the way for establishing a new method for definitive diagnosis.
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| Free Research Field |
分子生物学
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