Hyperactive transposase mutants of the Sleeping Beauty transposon
Hyperactive transposase mutants of the Sleeping Beauty transposon
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DOI：
10.1016/j.ymthe.2005.06.484
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发表时间：
2005-12-01
影响因子：
12.4
通讯作者：
Fletcher, BS
中科院分区：
文献类型：
--
作者：
Baus, J;Liu, L;Fletcher, BS
转座元件在克服非病毒基因传递的主要障碍之一，即缺乏长期基因表达方面具有巨大的潜力。睡美人(SB)转座子在哺乳动物细胞中具有最高的转座子活性，是一种很有前途的非病毒基因治疗载体系统。为了产生更有效的运输工具，我们对几个新的和以前发现的SB转座酶突变体进行了系统的评估。结果表明，某些突变体组合不能促进转座，而另一些突变体组合则产生协同反应。最活跃的突变体HSB17在相同的表达框中测试时，转座酶SB10的转座活性几乎是原始转座酶SB10的17倍。此外，当该超活性突变体与改进的转座子结合时，观察到协同活性。利用高活性转座酶的动物研究表明，报告基因的长期表达增强。这些修改进一步扩大了这种基于转座子的基因转移系统的用途。
Transposable elements have enormous potential to overcome one of the major hurdles in nonviral gene delivery, namely the lack of long-term gene expression. The Sleeping Beauty (SB) transposon is a promising vector system for nonviral gene therapy as it has the highest transposition activity of all known DNA transposons within mammalian cells. In an effort to generate a more efficient delivery vehicle, we conducted a systematic evaluation of several novel and previously identified SB transposase mutants. The results indicate that certain combinations of mutants do not enhance transposition, whereas others give a synergistic response. The most active mutant, designated HSB17, shows nearly 17-fold higher transposition activity compared to the original transposase SB10 when tested within the same expression cassette. In addition, synergistic activity is observed when this hyperactive mutant is combined with an improved transposon. Animal studies utilizing the hyperactive transposase show enhanced long-term reporter gene expression. These modifications further expand the utility of this transposon-based gene transfer system.