| RRC ID |
45480
|
| 著者 |
Yamamoto H, Ishimura M, Ochiai M, Takada H, Kusuhara K, Nakatsu Y, Tsuzuki T, Mitani K, Hara T.
|
| タイトル |
BTK gene targeting by homologous recombination using a helper-dependent adenovirus/adeno-associated virus hybrid vector.
|
| ジャーナル |
Gene Ther
|
| Abstract |
X-linked agammaglobulinemia (XLA) is one of the most common humoral immunodeficiencies, which is caused by mutations in Bruton's tyrosine kinase (BTK) gene. To examine the possibility of using gene therapy for XLA, we constructed a helper-dependent adenovirus/adeno-associated virus BTK targeting vector (HD-Ad.AAV BTK vector) composed of a genomic sequence containing BTK exons 6-19 and a green fluorescence protein-hygromycin cassette driven by a cytomegalovirus promoter. We first used NALM-6, a human male pre-B acute lymphoblastic leukemia cell line, as a recipient to measure the efficiency of gene targeting by homologous recombination. We identified 10 clones with the homologous recombination of the BTK gene among 107 hygromycin-resistant stable clones isolated from two independent experiments. We next used cord blood CD34⁺ cells as the recipient cells for the gene targeting. We isolated colonies grown in medium containing cytokines and hygromycin. We found that the targeting of the BTK gene occurred in four of the 755 hygromycin-resistant colonies. Importantly, the gene targeting was also observed in CD19⁺ lymphoid progenitor cells that were differentiated from the homologous recombinant CD34⁺ cells during growth in selection media. Our study shows the potential for the BTK gene therapy using the HD-Ad.AAV BTK vector via homologous recombination in hematopoietic stem cells.
|
| 巻・号 |
23(2)
|
| ページ |
205-13
|
| 公開日 |
2016-2-1
|
| DOI |
10.1038/gt.2015.91
|
| PII |
gt201591
|
| PMID |
26280081
|
| MeSH |
Agammaglobulinaemia Tyrosine Kinase
Agammaglobulinemia / genetics
Agammaglobulinemia / therapy
Cell Line, Tumor
Dependovirus / genetics*
Gene Targeting*
Genetic Diseases, X-Linked / genetics
Genetic Diseases, X-Linked / therapy
Genetic Therapy
Genetic Vectors / genetics*
Helper Viruses / genetics*
Homologous Recombination*
Humans
Male
Mutation
Protein-Tyrosine Kinases / genetics*
|
| IF |
4.128
|
| 引用数 |
7
|
|
WOS 分野
|
MEDICINE, RESEARCH & EXPERIMENTAL
BIOTECHNOLOGY & APPLIED MICROBIOLOGY
GENETICS & HEREDITY
BIOCHEMISTRY & MOLECULAR BIOLOGY
|
| リソース情報 |
| ヒト・動物細胞 |
NALM-6(RCB1933) |
