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Evolution of a T7 RNA Polymerase Variant That Transcribes 2' - O - Methyl RNA
Ellington, Andrew D.
Article from Journal - ilmiah internasional
Nature Biotechnology: The Science and Business of Biotechnology vol. 22 no. 9 (Sep. 2004)
Perpustakaan Pusat (Semanggi)
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Lihat Detail Induk
Modified RNA and DNA molecules have novel properties that their natural counterparts do not possess, such as better resistance to degredation in cells and improved pharmacokinetic behavior. In particular, modifications at the 2'-OH of ribose are important for enhancing the stability of RNA. Unfortunately, it is difficult to enzymatically synthesize modified nucleid acids of any substantial length because natural polymerases incorporate modified nucleotides inefficiently. Previously, we reported an activity based method for selecting functional T7 RNA polymerase to reproduce itself. Here we have modified the original procedure to identify polymerases that can efficiently incorporate multiple modified nucleotides at the 2' position of the ribose. Most important, our method allows the selection of polymerases that have good processivities and can be combined to simultaneously incorporate several different modified nucleotides in a unscript.
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