BTK gene targeting in a hematopoietic stem cell using an adenovirus adeno-associate virus hybrid vector

为了基因治疗x连锁无球蛋白血症，构建了含有布鲁顿酪氨酸激酶基因的辅助腺病毒腺相关病毒杂交载体。我们分析了通过同源重组（HR）进入造血干细胞（hsc）的靶向整合或基因修复的效率。人力资源效率达到0。每感染Nalm-6细胞073%在造血干细胞来源的人脐带血中，在3时检测到整合。每个扩散细胞6^* 10e-6， HR显示为0。7%的整合。此外，通过PCR检测DNA水平上的整合或HR，发现造血干细胞诱导的IgM阳性未成熟B细胞。我们还通过LAM-PCR表征了载体倾向于以非随机方式整合的染色体位点。

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.