Improved targeting of miRNA with antisense oligonucleotides.
MicroRNAs (miRNAs) are a class of 20-24 nt noncoding RNAs that regulate target mRNAs post-transcriptionally by binding with imperfect complementarity in the 3'-untranslated region (3'-UTR) and inhibiting translation or RNA stability. Current understanding of miRNA biology is limited, and antisense oligonucleotide (ASO) inhibition is a powerful technique for miRNA functionalization in vitro and in vivo, and for therapeutic targeting of miRNAs. Identification of optimal ASO chemistries for targeting miRNAs is therefore of great interest. We evaluated a number of 2'-sugar and backbone ASO modifications for their ability to inhibit miR-21 activity on a luciferase reporter mRNA. ASO modifications that improved target affinity improved miRNA ASO activity, yet the positioning of high-affinity modifications also had dramatically different effects on miRNA activity, suggesting that more than affinity determined the effectiveness of the miRNA ASOs. We present data in which the activity of a modified miRNA ASO was inversely correlated to its tolerability as an siRNA passenger strand, suggesting that a similar mechanism could be involved in the dissociation of miRNA ASOs and siRNA passenger strands. These studies begin to define the factors important for designing improved miRNA ASOs, enabling more effective miRNA functionalization and therapeutic targeting.
Davis
Scott
S
Isis Pharmaceuticals 1896 Rutherford Road, Carlsbad, CA 92008, USA.
Lollo
Bridget
B
Freier
Susan
S
Esau
Christine
C
D020137:Base Pair Mismatch; D002241:Carbohydrates / Q000737:chemistry; D017930:Genes, Reporter; D006367:HeLa Cells; D006801:Humans; D007700:Kinetics; D035683:MicroRNAs / Q000032:analysis / Q000037:antagonists & inhibitors* / Q000737:chemistry; D016376:Oligonucleotides, Antisense / Q000737:chemistry*; D010710:Phosphates / Q000737:chemistry; D034622:RNA Interference
D002241:Carbohydrates; D035683:MicroRNAs; D016376:Oligonucleotides, Antisense; D010710:Phosphates