Antisense oligonucleotide-directed cleavage of mRNA in Xenopus oocytes and eggs

Abstract

We have investigated the effect of specific antisense oligonucleotides on both exogenous and endogenous mRNAs in Xenopus oocytes and eggs. Injection of 19- or 20-mers complementary to 70-kd heat shock protein, histone H4 and vegetally localized Veg 1 coding sequences causes rapid cleavage and degradation of up to 96% of the target transcripts present in stage VI oocytes. Nuclear and cytoplasmic transcripts appear to be equally accessible to cytoplasmically injected oligonucleotide and efficient cleavage also occurs in mature oocytes and unfertilized eggs. The residual intact mRNA appears to be completely inaccessible, resisting cleavage by further addition of oligonucleotide. We confirm that antisense oligonucleotides appear to act specifically in vivo, as previously reported in vitro, by directing RNase H cleavage and destabilization of their complementary mRNA.