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This invention presents a method termed SELection of Modified Aptamers, or SELMA, allowing for the incorporation of large modifications into DNA libraries
About
By utilizing physical attachment of folded RNA libraries to their encoding DNA, this invention is presented as a way to circumvent the reverse transcription step during systematic evolution of RNA ligands by exponential enrichment (RNA‐SELEX). This method circumvents the need for reverse transcription in the amplification of RNA libraries and could be applied to base modifications for which reverse transcription is inefficient. The invention shows a verified method for DNA display of RNA. The benefits of using RNA in selections (structural diversity, amenability to 2’ modifications for nuclease resistance) can be coupled with substantial post-transcriptional modification in a SELMA-type experiment. Stringency was then increased by lowering the thrombin concentration from 10 to 1 nM and shortening incubation time from 1 hour to 5 minutes; after the 10th round of selection the library was cloned and 10 members sequenced and analyzed for thrombin binding.
Key Benefits
•An innovation for DNA display of RNA to circumvent the reverse transcription step during RNA-SELEX •Effectively displayed RNA library that could bind to human thrombin, with preferred secondary structures •A successful RNA library generation strategy allows the selected aptamers adopt a preferred conformation •Selection cycle is fast and straightforward since the display method uses commercially available materials