A cytoplasmic pathway for gapmer antisense oligonucleotide-mediated gene silencing in mammalian cells

Antisense oligonucleotides (ASOs) are known to trigger mRNA degradation in the nucleus via an RNase H-dependent mechanism. We have now identified a putative cytoplasmic mechanism through which ASO gapmers silence their targets when transfected or delivered gymnotically (i.e. in the absence of any tr...

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Bibliographic Details
Published in:Nucleic acids research 2015-10, Vol.43 (19), p.9350-9361
Main Authors: Castanotto, Daniela, Lin, Min, Kowolik, Claudia, Wang, LiAnn, Ren, Xiao-Qin, Soifer, Harris S, Koch, Troels, Hansen, Bo Rode, Oerum, Henrik, Armstrong, Brian, Wang, Zhigang, Bauer, Paul, Rossi, John, Stein, C A
Format: Article
Language:English
Subjects:
Argonaute Proteins - metabolism
Cell Line
Cytoplasm - chemistry
Cytoplasm - metabolism
Gene Silencing
Gene Transfer Techniques
Molecular Biology
Oligonucleotides, Antisense - analysis
Oligonucleotides, Antisense - chemistry
Oligonucleotides, Antisense - metabolism
RNA, Messenger - metabolism
RNA, Small Interfering
Transfection
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Summary:Antisense oligonucleotides (ASOs) are known to trigger mRNA degradation in the nucleus via an RNase H-dependent mechanism. We have now identified a putative cytoplasmic mechanism through which ASO gapmers silence their targets when transfected or delivered gymnotically (i.e. in the absence of any transfection reagent). We have shown that the ASO gapmers can interact with the Ago-2 PAZ domain and can localize into GW-182 mRNA-degradation bodies (GW-bodies). The degradation products of the targeted mRNA, however, are not generated by Ago-2-directed cleavage. The apparent identification of a cytoplasmic pathway complements the previously known nuclear activity of ASOs and concurrently suggests that nuclear localization is not an absolute requirement for gene silencing.
ISSN:0305-1048
1362-4962
DOI:10.1093/nar/gkv964