2011 Fiscal Year Final Research Report
BTK gene targeting in a hematopoietic stem cell using an adenovirus adeno-associate virus hybrid vector
| Project/Area Number |
21790996
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| Research Category |
Grant-in-Aid for Young Scientists (B)
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| Allocation Type | Single-year Grants |
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| Research Field |
Pediatrics
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| Research Institution | Kyushu University |
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Principal Investigator |
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| Research Collaborator |
ISHIMURA Masataka 九州大学, 医学研究院, 大学院生
YAMAMOTO Hiroyuki 九州大学, 医学研究院, 大学院生
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| Project Period (FY) |
2009 – 2011
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| Keywords | 移植 / 再生医療 / 遺伝子 / ウイルス |
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| Research Abstract |
For gene therapy of X-linked agammaglobulinemia, a helper-dependent adenovirus adeno-associated virus hybrid vector was constructed that contains Bruton' s tyrosin kinase gene. We analyzed an efficiency of targeted integration or gene repair through homologous recombination(HR) into hematopoietic stem cells(HSCs). The efficiency of HR reached 0. 073% per infected Nalm-6 cells. In HSCs derived human cord blood, integration was detected at 3. 6^* 10e-6 per spreading cells, and HR was revealed 0. 7% of those integrations. Furthermore, detection of integration or HR at DNA level by PCR revealed IgM positive-immature B cells that induced from HSCs. We also characterized chromosomal sites where the vector tends to integrate in a non-random manner by LAM-PCR.
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