PUBLICATION
Use of fully modified 2'-O-methyl antisense oligos for loss-of-function studies in vertebrate embryos
- Authors
- Schneider, P.N., Olthoff, J.T., Matthews, A.J., and Houston, D.W.
- ID
- ZDB-PUB-101115-18
- Date
- 2011
- Source
- Genesis (New York, N.Y. : 2000) 49(3): 117-123 (Journal)
- Registered Authors
- Keywords
- antisense oligonucleotides, embryo, 2'-O-methyl RNA, morpholino
- MeSH Terms
-
- Animals
- Bone Morphogenetic Protein 4/genetics
- Bone Morphogenetic Protein 4/metabolism
- Gene Expression Regulation, Developmental/drug effects
- Glycoproteins/genetics
- Glycoproteins/metabolism
- Intercellular Signaling Peptides and Proteins/genetics
- Intercellular Signaling Peptides and Proteins/metabolism
- Morpholines
- Morpholinos
- Oligoribonucleotides, Antisense/genetics*
- Oligoribonucleotides, Antisense/pharmacology
- Phenotype
- RNA Splicing
- RNA, Messenger
- Wnt Proteins/genetics
- Wnt Proteins/metabolism
- Xenopus Proteins/genetics
- Xenopus Proteins/metabolism
- Xenopus laevis/embryology*
- Xenopus laevis/genetics*
- Xenopus laevis/metabolism
- Zebrafish/embryology*
- Zebrafish/genetics*
- Zebrafish/metabolism
- beta Catenin/genetics
- beta Catenin/metabolism
- PubMed
- 21442720 Full text @ Genesis
Citation
Schneider, P.N., Olthoff, J.T., Matthews, A.J., and Houston, D.W. (2011) Use of fully modified 2'-O-methyl antisense oligos for loss-of-function studies in vertebrate embryos. Genesis (New York, N.Y. : 2000). 49(3):117-123.
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) 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) can efficiently and specifically generate embryonic loss-of-function phenotypes comparable to 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.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping