eSHAPE single RNA
In vitro secondary structure of single RNA molecules
eSHAPE single RNA is a method to obtain RNA structure probing data for in vitro transcribed RNA formulations. In vitro transcribed or synthetic RNAs are used as input for RNA structure probing with the NAI reagent. eSHAPE single RNA is available as a kit for end user completion, and as a full service offering with results delivered as a data package containing RNA structure information as reactivity values.
Selective 2’-Hydroxyl Acylation analyzed by Primer Extension (SHAPE) is a chemical probing method that measures RNA flexibility at single nucleotide resolution. As the exclusive licensee of the patented technology, we apply NAI to the RNA probing reaction and forms adducts with the free 2′-OH on the RNA backbone in single stranded regions. During the reverse transcription reaction these adducts induce mutations in the newly formed cDNA at a higher rate than the DMSO control sample. Mutation rates are then calculated for each position along the IVT RNA, and later computed into SHAPE reactivities that guide the folding of the RNA by indicating the pairing status for each position. The SHAPE technology on IVT-mRNAs measures local RNA flexibility in purified, deproteinized RNA in vitro, which can aid in the prediction of RNA secondary structure for the mRNA formulation.