Enzymatic combinatorial nucleoside deletion scanning mutagenesis of functional RNA

Abstract

We describe a general and simple method to identify catalytically and structurally important nucleotides in functional RNAs. Our approach is based on statistical replacement of each nucleoside with a non-nucleosidic spacer (C3 linker, Δ), followed by separation of active library variants and readout of interference effects by analysis of enzymatic primer extension reactions.