Use of fully modified 2'-O-methyl antisense oligos for loss-of-function studies in vertebrate embryos

Patricia N Schneider; John T Olthoff; Abby J Matthews; Douglas W Houston

doi:10.1002/dvg.20689

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Use of fully modified 2'-O-methyl antisense oligos for loss-of-function studies in vertebrate embryos

Journal article

Open access

Peer reviewed

Use of fully modified 2'-O-methyl antisense oligos for loss-of-function studies in vertebrate embryos

Patricia N Schneider, John T Olthoff, Abby J Matthews and Douglas W Houston

Genesis (New York, N.Y. : 2000), Vol.49(3), pp.117-123

03/2011

DOI: 10.1002/dvg.20689

PMCID: PMC3121920

PMID: 21442720

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Abstract

Antisense oligonucleotides are commonly employed to study the roles of genes in development. Although morpholino phosphorodiamidate oligonucleotides (morpholinos) are widely used to block translation or splicing of target gene products' the usefulness of other modifications in mediating RNase-H independent inhibition of gene activity in embryos has not been investigated. In this study, we investigated the extent that fully modified 2'-O-methyl oligonucleotides (2'-OMe oligos) that can function as translation inhibiting reagents in vivo, using Xenopus and zebrafish embryos. We find that oligos against Xenopus β-catenin, wnt11, and bmp4 and against zebrafish chordin (chd), which can efficiently and specifically generate embryonic loss-of-function phenotypes comparable with morpholino injection and other methods. These results show that fully modified 2'-OMe oligos can function as RNase-H independent antisense reagents in vertebrate embryos and can thus serve as an alternative modification to morpholinos in some cases.

Xenopus Proteins - genetics

Bone Morphogenetic Protein 4 - genetics

Glycoproteins - metabolism

Oligoribonucleotides, Antisense - pharmacology

Wnt Proteins - metabolism

Zebrafish - embryology

Bone Morphogenetic Protein 4 - metabolism

Intercellular Signaling Peptides and Proteins - metabolism

RNA Splicing

Wnt Proteins - genetics

Xenopus laevis - genetics

Glycoproteins - genetics

Xenopus laevis - embryology

Gene Expression Regulation, Developmental - drug effects

Intercellular Signaling Peptides and Proteins - genetics

Morpholines

beta Catenin - metabolism

Zebrafish - genetics

beta Catenin - genetics

Phenotype

Animals

Xenopus laevis - metabolism

Oligoribonucleotides, Antisense - genetics

Zebrafish - metabolism

Xenopus Proteins - metabolism

Morpholinos

RNA, Messenger

Details

Title: Subtitle: Use of fully modified 2'-O-methyl antisense oligos for loss-of-function studies in vertebrate embryos
Creators: Patricia N Schneider - Department of Biology, The University of Iowa, Iowa City, Iowa 52242-1324, USA
John T Olthoff
Abby J Matthews
Douglas W Houston
Resource Type: Journal article
Publication Details: Genesis (New York, N.Y. : 2000), Vol.49(3), pp.117-123
DOI: 10.1002/dvg.20689
PMID: 21442720
PMCID: PMC3121920
NLM abbreviation: Genesis
ISSN: 1526-954X
eISSN: 1526-968X
Publisher: United States
Grant note: R01 GM083999 / NIGMS NIH HHS 5R01GM083999-02 / NIGMS NIH HHS R01 GM083999-05 / NIGMS NIH HHS R01 GM083999-03 / NIGMS NIH HHS
Language: English
Date published: 03/2011
Academic Unit: Biology
Record Identifier: 9983992045802771

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