Activities of functional RNA molecules containing unnatural bases in cultured cells

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By using in vitro transcription with our unnatural base pair systems, we prepared shRNA molecules containing a series of unnatural bases at a specific position and evaluated their RNAi activities in cultured cells. We found that the activity was retained by replacement with unnatural bases in some specific positions within the passenger and guide strands of the shRNA molecules, and that the tolerance by unnatural-base replacements depended on the size of the unnatural base.