| Project/Area Number |
26670505
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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 |
Pediatrics
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| Research Institution | Hamamatsu University School of Medicine (2015)
Yokohama City University (2014) |
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Principal Investigator |
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| Project Period (FY) |
2014-04-01 – 2016-03-31
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| Project Status |
Completed (Fiscal Year 2015)
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| Budget Amount *help |
¥3,640,000 (Direct Cost: ¥2,800,000、Indirect Cost: ¥840,000)
Fiscal Year 2015: ¥1,040,000 (Direct Cost: ¥800,000、Indirect Cost: ¥240,000)
Fiscal Year 2014: ¥2,600,000 (Direct Cost: ¥2,000,000、Indirect Cost: ¥600,000)
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| Keywords | スプライス変異 / ターゲットキャプチャ－ / 次世代シークエンス / スプライシング / RNA / 次世代シークエンサー / 遺伝学 |
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| Outline of Final Research Achievements |
Lymphoblastoid cells derived from seven patients with mutations causing abnormal splicing and a patient with a possible mutation only in one allele were examined by target RNA capture against 24 genes. Enriched libraries were sequenced in one lane of HiSeq 2500 sequencer, and reads were aligned by Novoalign to transcriptome or by TopHat to reference genome. Although Novoalign was able to align more reads with exon skipping, it failed to align reads with intron retention. On the other hand, TopHat was able to align both reads with exon skipping and intron retention, suggesting that TopHat is first choice for alignment. In lymphoblastoid cells, five out of 24 genes are not well sequenced, indicating that even in target capture system, tissues responsible for disease are required for optimized analysis.
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