Retroviral vectors aimed at the gene therapy of human beta-globin gene disorders.
Retroviral vectors aimed at the gene therapy of human beta-globin gene disorders.
复制标题
逆转录病毒载体旨在人类β-珠蛋白基因疾病的基因治疗。
DOI：
10.1111/j.1749-6632.1998.tb10472.x
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发表时间：
1998
影响因子：
5.2
通讯作者：
Leboulch,P
中科院分区：
文献类型：
--
作者：
Pawliuk,R;Bachelot,T;Raftopoulos,H;Kalberer,C;Humphries,RK;Bank,A;Leboulch,P
翻译后摘要：我们正专注于开发复杂的逆转录病毒载体含有人类β珠蛋白基因和β LCR的基因治疗镰状细胞病和β地中海贫血。第一代载体含有突变的剪接位点，以确保前病毒转移的稳定性，使10/12只移植小鼠能够长期重建至少8个月，分析的3只小鼠中有2只具有高表达水平（5%和20%鼠β）。在第二受体中也实现了转移和表达（范围：3-11%鼠β）。未观察到位置独立表达。为了提高基因转移的效率，并获得受体小鼠与专门转导的细胞的完全重建，同时富集前病毒整合到活性染色质区域中，我们掺入了表达CD 24或绿色荧光蛋白（GFP）的盒。稳定转移至鼠骨髓细胞允许对转导细胞的纯群体进行有效的FACS分选。基于这些原理并含有γ-或δ-珠蛋白基因片段的载体家族也被设计用于系统分析其抗镰状化特性。
Abstract:We are focusing on the development of complex retroviral vectors containing human β‐globin gene and β‐LCR for the gene therapy of sickle cell disease and β‐thalassemias. First generation vectors containing mutated splice‐sites to insure stability of proviral transfer enabled long‐term reconstitution in 10/12 transplanted mice for a least 8 months with high expression levels in 2 out of 3 mice analyzed (5% and 20% murine β). Transfer and expression were also achieved in secondary recipients (range: 3–11% murine β). Position independent expression was not observed. In an effort to increase the efficiency of gene transfer and obtain complete reconstitution of recipient mice with exclusively transduced cells while enriching for proviral integration into active chromatin regions, we have incorporated a cassette expressing CD24 or the green fluorescent protein (GFP). Stable transfer to murine bone marrow cells allowed efficient FACS‐sorting of pure populations of transduced cells. A family of vectors based on these principles and containing segments of γ‐ or δ‐globin genes were also designed for systematic analysis of their anti‐sickling properties.