Silencing of microRNA families by seed-targeting tiny LNAs

被引：496

作者：

Obad, Susanna ^{[1
]}

dos Santos, Camila O. ^{[2
]}

Petri, Andreas ^{[1
]}

Heidenblad, Markus ^{[1
]}

Broom, Oliver ^{[1
]}

Ruse, Cristian ^{[2
]}

Fu, Cexiong ^{[2
]}

Lindow, Morten ^{[1
]}

Stenvang, Jan ^{[1
]}

Straarup, Ellen Marie ^{[1
]}

Hansen, Henrik Frydenlund ^{[1
]}

Koch, Troels ^{[1
]}

Pappin, Darryl ^{[2
]}

Hannon, Gregory J. ^{[2
]}

Kauppinen, Sakari ^{[1
,3
]}

机构：

[1] Santaris Pharma, Horsholm, Denmark

[2] Cold Spring Harbor Lab, Cold Spring Harbor, NY USA

[3] Aalborg Univ, Copenhagen Inst Technol, Ballerup, Denmark

来源：

NATURE GENETICS | 2011年 / 43卷 / 04期

基金：

英国医学研究理事会;

关键词：

LOCKED NUCLEIC-ACID; IN-VIVO; ANTISENSE OLIGONUCLEOTIDES; MESSENGER-RNA; LET-7; EXPRESSION; MIR-21; MICE; DETERMINANTS; SPECIFICITY;

D O I：

10.1038/ng.786

中图分类号：

Q3 [遗传学];

学科分类号：

071007 ; 090102 ;

摘要：

The challenge of understanding the widespread biological roles of animal microRNAs (miRNAs) has prompted the development of genetic and functional genomics technologies for miRNA loss-of-function studies. However, tools for exploring the functions of entire miRNA families are still limited. We developed a method that enables antagonism of miRNA function using seed-targeting 8-mer locked nucleic acid (LNA) oligonucleotides, termed tiny LNAs. Transfection of tiny LNAs into cells resulted in simultaneous inhibition of miRNAs within families sharing the same seed with concomitant upregulation of direct targets. In addition, systemically delivered, unconjugated tiny LNAs showed uptake in many normal tissues and in breast tumors in mice, coinciding with long-term miRNA silencing. Transcriptional and proteomic profiling suggested that tiny LNAs have negligible off-target effects, not significantly altering the output from mRNAs with perfect tiny LNA complementary sites. Considered together, these data support the utility of tiny LNAs in elucidating the functions of miRNA families in vivo.

引用

页码：371 / U160

页数：10

共 54 条

[1] The let-7 microRNA family members mir-48, mir-84, and mir-241 function together to regulate developmental timing in Caenorhabditis elegans [J].