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EC Number Application Commentary Reference
Display the word mapDisplay the reaction diagram Show all sequences 4.6.1.16molecular biology controlled RNA splicing in mammalian cells. mRNA modification technology that makes use of tRNA splicing endonuclease and its natural substrate, the bulge-helix-bulge structure. These components can perform both cis- and trans-splicing in cellular and animal models and may provide a convenient way to modulate gene expression using components independent of cellular regulatory networks. To use the Methanocaldococcus jannaschii enzyme in stable expression mammalian systems, variants are developed which are characterized by high efficiency and sustainable in vivo activity. The variants are created by the introduction of proper localization signals followed by mutagenesis and direct selection in mammalian cells. The best endonuclease variant shows 40fold higher activity compared to the parental enzyme and stable processing of 30% of the target mRNA. These variants show complete compatibility with long-term expression in mammalian cells, suggesting that they may be usefully applied in functional genomics and genetically modified animal models -, 727463
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