Handling S/MAR vectors.
Handling S/MAR vectors.
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DOI：
10.1101/pdb.top068262
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发表时间：
2012-06-01
影响因子：
--
通讯作者：
Lipps, Hans J
中科院分区：
文献类型：
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作者：
Hagedorn, Claudia;Baiker, Armin;Lipps, Hans J
非病毒附加型载体代表了当前使用的基于病毒的表达系统的有吸引力的替代品。 20 世纪 90 年代末，研究表明，含有与支架/基质附着区 (S/MAR) 连接的表达盒的质粒在所有测试的细胞系以及原代细胞中以低拷贝数附加体的形式复制，并且可用于高等动物的遗传修饰。一旦在细胞中建立，S/MAR 载体就会在 S 期早期复制，并且在没有选择的情况下，可以无限期地稳定保留在细胞中。因此，该载体可以被视为研究复制和功能性核结构的表观遗传调控的最小模型系统。理论上，这种构建体代表了基因治疗几乎“理想”的表达系统。实际上，基于 S/MAR 的载体可以稳定地修饰哺乳动物细胞，其效率远低于基于病毒的构建体。因此，它们尚未在基因治疗试验中得到应用。此外，S/MAR 载体系统处理起来并不简单，在针对各种应用修改这些载体时必须考虑几个关键的技术问题。
Nonviral episomal vectors represent attractive alternatives to currently used virus-based expression systems. In the late 1990s, it was shown that a plasmid containing an expression cassette linked to a scaffold/matrix attached region (S/MAR) replicates as a low copy number episome in all cell lines tested, as well as primary cells, and can be used for the genetic modification of higher animals. Once established in the cell, the S/MAR vector replicates early during S-phase and, in the absence of selection, is stably retained in the cells for an unlimited period of time. This vector can therefore be regarded as a minimal model system for studying the epigenetic regulation of replication and functional nuclear architecture. In theory, this construct represents an almost "ideal" expression system for gene therapy. In practice, S/MAR-based vectors stably modify mammalian cells with efficiencies far below those of virus-based constructs. Consequently, they have not yet found application in gene therapy trials. Furthermore, S/MAR vector systems are not trivial to handle and several critical technical issues have to be considered when modifying these vectors for various applications.