Vika/vox, a novel efficient and specific Cre/loxP-like site-specific recombination system.
Vika/vox, a novel efficient and specific Cre/loxP-like site-specific recombination system.
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DOI：
10.1093/nar/gks1037
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发表时间：
2013-01
影响因子：
14.9
通讯作者：
Buchholz F
中科院分区：
文献类型：
--
作者：
Karimova M;Abi-Ghanem J;Berger N;Surendranath V;Pisabarro MT;Buchholz F
靶向基因组工程已成为不同学科的重要研究领域，其中位点特异性重组酶（SSR）是最流行的基因组工程工具之一。它们触发切除、整合、倒位和易位的能力使得SSR成为体外和体内操纵DNA的宝贵工具。然而，结合联合收割机不同SSR系统的复杂策略不断增加。因此，对额外的精确和有效的重组酶的需求是由遗传研究的日益复杂性决定的。在这里，我们描述了一种新的位点特异性重组系统Vika/VOX。Vika来源于溶珊瑚弧菌的退化噬菌体，与其他酪氨酸重组酶具有低序列相似性，但功能上进行类似类型的反应。我们证明，Vika是高度特异性的催化VOX重组，而不重组其他SSR系统的靶位点。我们还比较了Vika/Vox与其他SSR系统的重组活性，为决定最适合特定应用的酶提供了指导，并证明Vika表达不会引起哺乳动物细胞的细胞毒性。我们的研究结果表明，Vika/vox是一种新的强大和安全的工具，在'遗传工具箱'，可以单独使用或与其他SSR在异源宿主中组合。
Targeted genome engineering has become an important research area for diverse disciplines, with site-specific recombinases (SSRs) being among the most popular genome engineering tools. Their ability to trigger excision, integration, inversion and translocation has made SSRs an invaluable tool to manipulate DNA in vitro and in vivo. However, sophisticated strategies that combine different SSR systems are ever increasing. Hence, the demand for additional precise and efficient recombinases is dictated by the increasing complexity of the genetic studies. Here, we describe a novel site-specific recombination system designated Vika/vox. Vika originates from a degenerate bacteriophage of Vibrio coralliilyticus and shares low sequence similarity to other tyrosine recombinases, but functionally carries out a similar type of reaction. We demonstrate that Vika is highly specific in catalyzing vox recombination without recombining target sites from other SSR systems. We also compare the recombination activity of Vika/vox with other SSR systems, providing a guideline for deciding on the most suitable enzyme for a particular application and demonstrate that Vika expression does not cause cytotoxicity in mammalian cells. Our results show that Vika/vox is a novel powerful and safe instrument in the ‘genetic toolbox’ that can be used alone or in combination with other SSRs in heterologous hosts.
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影响因子：
5.3
作者：
Forni, Paolo E.;Scuoppo, Claudio;Ponzetto, Carola
通讯作者：
Ponzetto, Carola
影响因子：
2.7
作者：
Buchholz, F;Bishop, JM
通讯作者：
Bishop, JM
影响因子：
4.4
作者：
JORGENSEN, WL;CHANDRASEKHAR, J;KLEIN, ML
通讯作者：
KLEIN, ML
影响因子：
46.9
作者：
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通讯作者：
Stewart, AF
影响因子：
64.8
作者：
Guo, F;Gopaul, DN;VanDuyne, GD
通讯作者：
VanDuyne, GD