High-titre retroviral vector system for efficient gene delivery into human and mouse cells of haematopoietic and lymphocytic lineages.
High-titre retroviral vector system for efficient gene delivery into human and mouse cells of haematopoietic and lymphocytic lineages.
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DOI：
10.1099/vir.0.020255-0
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发表时间：
2010-08
期刊：
影响因子：
--
通讯作者：
Lu Y
中科院分区：
文献类型：
--
作者：
Wu C;Lu Y
造血和淋巴细胞谱系的基因修饰细胞可以为广泛的遗传和获得性疾病提供潜在的治愈性治疗。然而，由于慢病毒载体的低效率，这种应用在小鼠模型中受到限制。为了促进用于增强基因递送的高滴度无辅助病毒逆转录病毒载体的快速生产，对原型莫洛尼鼠白血病病毒（MoMLV）衍生的载体系统进行了多种修饰，包括使载体系统适应包装细胞中猿猴病毒40 ori/T抗原介导的附加型复制，用一系列更强的复合启动子替换MoMLV 5′ U3启动子，并在3′长末端重复序列下游添加额外的多聚腺苷酸化信号。这些修饰使载体生产提高了2-3个对数。测试高滴度载体储备液感染多种来源于人和小鼠的细胞的能力，包括原代单核细胞来源的巨噬细胞培养物。虽然慢病毒载体在整合水平上受到显著限制，但基于MoMLV的载体显示出小鼠细胞的有效基因转导。这种高滴度的逆转录病毒载体系统代表了一种有用的工具，用于有效地将基因递送到人类和小鼠的造血细胞和淋巴细胞中，特别是在小鼠中作为新的基因治疗试验的小动物模型。
Genetically modified cells of haematopoietic and lymphocytic lineages could provide potentially curative treatments for a wide range of inherited and acquired diseases. However, this application is limited in mouse models by the low efficiency of lentiviral vectors. To facilitate the rapid production of high-titre helper-free retroviral vectors for enhanced gene delivery, multiple modifications to a prototype moloney murine leukemia virus (MoMLV)-derived vector system were made including adaptation of the vector system to simian virus 40 ori/T antigen-mediated episomal replication in packaging cells, replacement of the MoMLV 5′ U3 promoter with a series of stronger composite promoters and addition of an extra polyadenylation signal downstream of the 3′ long terminal repeat. These modifications enhanced vector production by 2–3 logs. High-titre vector stocks were tested for their ability to infect a variety of cells derived from humans and mice, including primary monocyte-derived macrophage cultures. Whilst the lentiviral vector was significantly restricted at the integration level, the MoMLV-based vector showed effective gene transduction of mouse cells. This high-titre retroviral vector system represents a useful tool for efficient gene delivery into human and mouse haematopoietic and lymphocytic cells, with particular application in mice as a small animal model for novel gene therapy tests.
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影响因子：
3.6
作者：
Hawley, Robert G.
通讯作者：
Hawley, Robert G.
影响因子：
4.1
作者：
May, Tobias;Butueva, Milada;Wirth, Dagmar
通讯作者：
Wirth, Dagmar
DOI：
10.1073/pnas.232688199
发表时间：
2003-01-07
影响因子：
11.1
作者：
Qin, XF;An, DS;Baltimore, D
通讯作者：
Baltimore, D
DOI：
10.1073/pnas.90.18.8392
发表时间：
1993-09-15
影响因子：
11.1
作者：
PEAR, WS;NOLAN, GP;BALTIMORE, D
通讯作者：
BALTIMORE, D
影响因子：
168.9
作者：
Fischer, Alain;Cavazzana-Calvo, Marina
通讯作者：
Cavazzana-Calvo, Marina