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

为用于X连锁无丙种球蛋白血症的基因治疗，构建了含有Bruton‘S酪氨酸激酶基因的辅助子依赖型腺病毒腺相关病毒混合载体。我们分析了通过同源重组(HR)定向整合或基因修复进入造血干细胞(HSCs)的效率。人力资源效率达到0。感染NALM-6细胞的细胞百分率为073%。在人脐血来源的HSCs中，每个铺展细胞在3.6^*10e-6处检测到整合，HR为0。这些集成中的7%。此外，在DNA水平上检测整合或HR，发现从HSC中诱导出了IgM阳性的未成熟B细胞。我们还通过LAM-PCR对载体倾向于以非随机方式整合的染色体位置进行了表征。

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.