Vika/vox, a novel efficient and specific Cre/loxP-like site-specific recombination system

被引:55
作者
Karimova, Madina [1 ,2 ]
Abi-Ghanem, Josephine [3 ]
Berger, Nicolas [1 ,2 ]
Surendranath, Vineeth [4 ]
Pisabarro, Maria Teresa [3 ]
Buchholz, Frank [1 ,2 ]
机构
[1] Univ Technol, Fac Med, D-01307 Dresden, Germany
[2] Univ Technol, Univ Hosp Carl Gustav Carus, D-01307 Dresden, Germany
[3] BIOTEC TU Dresden, D-01037 Dresden, Germany
[4] Max Planck Inst Mol Cell Biol & Genet, D-01307 Dresden, Germany
关键词
CRE RECOMBINASE; FLP RECOMBINASE; DIRECTED EVOLUTION; DNA RECOMBINATION; GENOME; CELLS; RECOGNITION; EXPRESSION; MULTIPLE; VECTORS;
D O I
10.1093/nar/gks1037
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
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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页数:13
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