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

Adgang til dokumentet

10.1038/ng.786

Citationsformater

@article{fccad32101aa45f09951a43ff8f9ccec,

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.",

keywords = "3' Untranslated Regions, Animals, Base Sequence, Cell Line, Tumor, Female, Gene Knockdown Techniques, Gene Silencing, Genes, Reporter, Genetic Techniques, HeLa Cells, Humans, Liver, Luciferases, Renilla, Mammary Neoplasms, Experimental, Mice, Mice, Inbred BALB C, MicroRNAs, Oligonucleotides",

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

year = "2011",

month = feb,

doi = "10.1038/ng.786",

language = "English",

volume = "43",

pages = "371--8",

journal = "Nature: New biology",

issn = "1061-4036",

publisher = "nature publishing group",

number = "4",

}

TY - JOUR

T1 - Silencing of microRNA families by seed-targeting tiny LNAs

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

Y1 - 2011/2

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.

KW - 3' Untranslated Regions

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

M3 - Journal article

C2 - 21423181

SN - 1061-4036

VL - 43

SP - 371

EP - 378

JO - Nature: New biology

JF - Nature: New biology

IS - 4

ER -

Silencing of microRNA families by seed-targeting tiny LNAs

Abstract

Adgang til dokumentet

Fingeraftryk

Citationsformater