The Sleeping Beauty transposon toolbox.
The Sleeping Beauty transposon toolbox.
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DOI：
10.1007/978-1-61779-603-6_13
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发表时间：
2012-01-01
期刊：
影响因子：
--
通讯作者：
Ivics, Zoltan
中科院分区：
文献类型：
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作者：
Ammar, Ismahen;Izsvak, Zsuzsanna;Ivics, Zoltan
II 类转座元件（DNA 转座子）的移动性可以通过分离转座子的两个功能组件进行实验控制：位于待移动的感兴趣基因侧翼的末端反向重复序列和可以有条件提供以驱动转座反应的转座酶蛋白。因此，克隆在基于转座子的载体的反向重复序列之间的感兴趣的DNA分子（例如，荧光标记、shRNA表达盒、诱变基因捕获或治疗基因构建体）可以以受控且高效的方式稳定地整合到基因组中。睡美人 (SB) 是第一个能够在脊椎动物细胞中进行基因转移的转座子，最近的结果证实，SB 支持脊椎动物物种的全方位基因工程，包括转基因、插入诱变和治疗性体细胞基因，以及离体和体内转移。这种方法论范式为基础和应用研究的基因组操作开辟了许多途径。本综述重点介绍了 SB 转座子技术在多种遗传应用中的最新技术，特别重点介绍了 SB 转座子工具箱中目前可用的转座子和转座酶载体。
The mobility of class II transposable elements (DNA transposons) can be experimentally controlled by separating the two functional components of the transposon: the terminal inverted repeat sequences that flank a gene of interest to be mobilized and the transposase protein that can be conditionally supplied to drive the transposition reaction. Thus, a DNA molecule of interest (e.g., a fluorescent marker, an shRNA expression cassette, a mutagenic gene trap or a therapeutic gene construct) cloned between the inverted repeat sequences of a transposon-based vector can be stably integrated into the genome in a regulated and highly efficient manner. Sleeping Beauty (SB) was the first transposon ever shown capable of gene transfer in vertebrate cells, and recent results confirm that SB supports a full spectrum of genetic engineering in vertebrate species, including transgenesis, insertional mutagenesis, and therapeutic somatic gene, transfer both ex vivo and in vivo. This methodological paradigm opened up a number of avenues for genome manipulations for basic and applied research. This review highlights the state-of-the-art in SB transposon technology in diverse genetic applications with special emphasis on the transposon as well as transposase vectors currently available in the SB transposon toolbox.