2015 Fiscal Year Final Research Report
BTK gene repair by homologous recombination using a helper-dependent adenovirus/adeno-associated virus hybrid vector
| Project/Area Number |
26860811
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| Research Category |
Grant-in-Aid for Young Scientists (B)
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| Allocation Type | Multi-year Fund |
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| Research Field |
Pediatrics
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| Research Institution | Kyushu University |
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Principal Investigator |
YAMAMOTO Hiroyuki 九州大学, 環境発達医学研究センター, 学術研究員 (00710170)
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| Project Period (FY) |
2014-04-01 – 2016-03-31
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| Keywords | 遺伝子変異修復 |
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| Outline of Final Research Achievements |
We constructed a new helper-dependent adenovirus/adeno-associated virus hybrid vector (HD-Ad.AAV hybrid vector) that is composed of the genomic sequence containing BTK exons 2-19, and GFP/puromycin resistant gene. Using this HD-Ad.AAV hybrid vector and CRISPR-Cas9, we found that the targeting in the BTK gene occurred in 10 among 24 hygromycin-resistant BFU-E colonies differentiated from the cord blood CD34+ cells. The estimated targeting efficiency was 1.8 x10-5 per cell. Our study shows the possibility to perform gene repair of BTK gene by homologous recombination in hematopoietic stem cells using this HD-Ad.AAV hybrid vector and CRISPR-Cas9 system.
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| Free Research Field |
分子生物学
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