An LNA-based loss-of-function assay for micro-RNAs.
Biomed Pharmacother
; 60(9): 633-8, 2006 Nov.
Article
in En
| MEDLINE
| ID: mdl-16962735
ABSTRACT
MicroRNAs (miRNAs) have recently emerged as being essential for development and for the control of cell proliferation/differentiation in various organisms. However, little is known about miRNA function and mode of action at the cellular level. We have designed a miRNA loss-of-function assay, based on chemically modified locked nucleic acids (LNA) antisense oligonucleotides and usable in tissue culture cells. We show that LNA/DNA mixed oligonucleotides form highly stable duplexes with miRNAs in vitro. Ex vivo, the target miRNA becomes undetectable in cells transfected with the antisense oligonucleotide. The effect is dose-dependent, long-lasting, and specific. Moreover, using a reporter assay, we show that antisense LNA/DNA oligonucleotides inhibit short non-coding RNAs at the functional level. Thus LNA/DNA mixmers represent powerful tools for functional analysis of miRNAs.
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Collection:
01-internacional
Database:
MEDLINE
Main subject:
Oligonucleotides, Antisense
/
MicroRNAs
Limits:
Humans
Language:
En
Journal:
Biomed Pharmacother
Year:
2006
Type:
Article
Affiliation country:
France