Quantification of Oligonucleotide Association with miRNA-Argonaute Complexes In Vitro

A major challenge in the development of oligonucleotide-based microRNA (miRNA) inhibitors for therapeutic applications is the identification of candidate designs with strong affinity for the target miRNA in the context of the Argonaute complex. To this effect, distinct chemical modifications are employed along the length of the oligonucleotide aimed at strengthening the interactions with the target miRNA. However, the modification chemistry and placement can inadvertently affect the intrinsic ability of the oligonucleotide to pair with its target in the context of Argonaute. To facilitate the design of potent oligonucleotides, we developed a sensitive high-throughput methodology to compare anti-miR compounds for their ability to associate with the miRNA/Argonaute complex.