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Genesis March 1, 2011; 49 (3): 117-23.

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

Schneider PN , Olthoff JT , Matthews AJ , Houston DW .

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.

PubMed ID: 21442720
PMC ID: PMC3121920
Article link: Genesis
Grant support: [+]

Species referenced: Xenopus laevis
Genes referenced: bmp4 cat.2 chrd.1 ctnnb1 gsc nodal3.1 otx2 pax8 sia1 szl wnt11
Morpholinos: bmp4 MO1 ctnnb1 MO1 wnt11b MO1

Article Images: [+] show captions
References [+] :
Boiziau, Inhibition of translation initiation by antisense oligonucleotides via an RNase-H independent mechanism. 1991, Pubmed, Xenbase