Single-stranded microRNA mimics

miRNAs are ∼22-nt RNAs that bind to the Argonaute family of proteins and have important regulatory roles in plants and animals. Here, we show that miRNAs exhibit targeting activity in cells when delivered as single strands that are 5'-phosphorylated and that contain 2'-fluoro ribose modifi...

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Bibliographic Details
Published in:RNA (Cambridge) 2012-10, Vol.18 (10), p.1796-1804
Main Authors: Chorn, Guillaume, Klein-McDowell, Molly, Zhao, Lihong, Saunders, Matthew A, Flanagan, W Michael, Willingham, Aarron T, Lim, Lee P
Format: Article
Language:English
Subjects:
Base Sequence
Cloning, Molecular
DNA, Single-Stranded - chemical synthesis
DNA, Single-Stranded - chemistry
Fluorides - chemical synthesis
Fluorides - chemistry
Gene Knockdown Techniques - instrumentation
Gene Knockdown Techniques - methods
HCT116 Cells
High-Throughput Screening Assays
Humans
Microarray Analysis
MicroRNAs - chemistry
Molecular Mimicry - physiology
Molecular Sequence Data
Phosphorylation
Ribose - chemical synthesis
Ribose - chemistry
Transfection
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Summary:miRNAs are ∼22-nt RNAs that bind to the Argonaute family of proteins and have important regulatory roles in plants and animals. Here, we show that miRNAs exhibit targeting activity in cells when delivered as single strands that are 5'-phosphorylated and that contain 2'-fluoro ribose modifications. Length preferences, chemical modification sensitivity, and genome-wide seed-based targeting all suggest that this activity is Ago-based. Activity could be enhanced by annealing of segmented passenger strands containing non-nucleic acid spacers. Furthermore, screening of randomly generated sequences identified pyrimidine rich 3' cassette sequences that increased single strand activity. These results provide an initial step in the development of single-stranded miRNA mimics for therapeutic use.
ISSN:1355-8382
1469-9001
DOI:10.1261/rna.031278.111