Antisense inhibition of beta-glucuronidase expression in preimplantation mouse embryos: a comparison of transgenes and oligodeoxynucleotides.
Using as a model the inhibition of beta-glucuronidase expression in preimplantation embryos, we have compared injections of transgenes directing the synthesis of antisense RNA and antisense oligodeoxynucleotides to our previous results with cytoplasmic injections of antisense RNAs. Pronuclear injection of an antisense DNA construct containing 1.4 kb of coding region of beta-glucuronidase fused to the mouse metallothionein I promoter results in transient inhibition of gene expression in preimplantation mouse embryos. Pronuclear injection of a smaller antisense DNA construct, overlapping the start codon, failed to inhibit gene expression. Injection of two 20-mer antisense oligodeoxynucleotides, one complementary to sequences including the initiation codon and the second complementary to exon 7 sequences of the beta-glucuronidase gene, failed to inhibit gene expression. In addition, cultures of embryos in the presence of antisense oligodeoxynucleotides had no effect on gene activity. Using radiolabeled oligomers added to the culture medium, we found poor uptake of oligodeoxynucleotides by embryos.
Ao
A
A
Department of Human Genetics, University of Michigan School of Medicine, Ann Arbor 48109.
Erickson
R P
RP
Bevilacqua
A
A
Karolyi
J
J
D000818:Animals; D001483:Base Sequence; D001755:Blastocyst / Q000187:drug effects / Q000201:enzymology / Q000378:metabolism*; D046508:Culture Techniques; D003592:Cytophotometry; D016373:DNA, Antisense / Q000235:genetics / Q000494:pharmacology*; D015870:Gene Expression / Q000187:drug effects*; D005966:Glucuronidase / Q000096:biosynthesis* / Q000235:genetics; D007267:Injections; D051379:Mice; D008810:Mice, Inbred C57BL; D008969:Molecular Sequence Data; D016376:Oligonucleotides, Antisense / Q000235:genetics / Q000494:pharmacology*
D016373:DNA, Antisense; D016376:Oligonucleotides, Antisense; D005966:Glucuronidase