Silencing of microRNA families by seed-targeting tiny LNAs

Susanna Obad; Camila O dos Santos; Andreas Petri; Markus Heidenblad; Oliver Broom; Cristian Ruse; Cexiong Fu; Morten Lindow; Jan Stenvang; Ellen Marie Straarup; Henrik Frydenlund Hansen; Troels Koch; Darryl Pappin; Gregory J Hannon; Sakari Kauppinen

doi:10.1038/ng.786

Silencing of microRNA families by seed-targeting tiny LNAs

Susanna Obad, Camila O dos Santos, Andreas Petri, Markus Heidenblad, Oliver Broom, Cristian Ruse, Cexiong Fu, Morten Lindow, Jan Stenvang, Ellen Marie Straarup, Henrik Frydenlund Hansen, Troels Koch, Darryl Pappin, Gregory J Hannon, Sakari Kauppinen

492 Citationer (Scopus)

Abstract

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.

Originalsprog	Engelsk
Tidsskrift	Nature Genetics
Vol/bind	43
Udgave nummer	4
Sider (fra-til)	371-8
Antal sider	8
DOI	https://doi.org/10.1038/ng.786
Status	Udgivet - feb. 2011
Udgivet eksternt	Ja

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10.1038/ng.786

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title = "Silencing of microRNA families by seed-targeting tiny LNAs",

abstract = "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.",

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doi = "10.1038/ng.786",

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AU - Obad, Susanna

AU - dos Santos, Camila O

AU - Petri, Andreas

AU - Heidenblad, Markus

AU - Broom, Oliver

AU - Ruse, Cristian

AU - Fu, Cexiong

AU - Lindow, Morten

AU - Stenvang, Jan

AU - Straarup, Ellen Marie

AU - Hansen, Henrik Frydenlund

AU - Koch, Troels

AU - Pappin, Darryl

AU - Hannon, Gregory J

AU - Kauppinen, Sakari

PY - 2011/2

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N2 - 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.

AB - 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.

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KW - Animals

KW - Base Sequence

KW - Cell Line, Tumor

KW - Female

KW - Gene Knockdown Techniques

KW - Gene Silencing

KW - Genes, Reporter

KW - Genetic Techniques

KW - HeLa Cells

KW - Humans

KW - Liver

KW - Luciferases, Renilla

KW - Mammary Neoplasms, Experimental

KW - Mice

KW - Mice, Inbred BALB C

KW - MicroRNAs

KW - Oligonucleotides

U2 - 10.1038/ng.786

DO - 10.1038/ng.786

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VL - 43

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JO - Nature Genetics

JF - Nature Genetics

IS - 4

ER -

Silencing of microRNA families by seed-targeting tiny LNAs

Abstract

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