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Nucleic Acids Res May 15, 2000; 28 (10): 2153-7.

Targeted elimination of zygotic messages in Xenopus laevis embryos by modified oligonucleotides possessing terminal cationic linkages.

Dagle JM , Littig JL , Sutherland LB , Weeks DL .

We have designed a new class of modified antisense oligodeoxyribonucleotides (ODN) consisting of a central contiguous stretch of 6-8 unmodified nucleotides flanked by 3''- and 5''-regions containing several nucleotides joined by cationic internucleoside linkages. The positive charge results from modification of the internucleoside linkages as N, N -diethylethylene-diamine phosphoramidates. These zwitterionic compounds show improved antisense activity in both Xenopus oocytes and embryos compared to our previously described chimeric oligonucleotides possessing neutral terminal internucleoside linkages. Using the localized maternal mRNA An2 as a target, we have shown that chimeric oligonucleotides with terminal positive charges are very effective in the sequence-specific elimination of maternal messages present in both oocytes and embryos. In addition, using the embryonic mRNA GS17 as a target, we have shown that these oligonucleotides can direct RNase H-mediated cleavage of messages produced at the onset of zygotic transcription, after the mid-blastula stage. These new compounds should be useful in attenuating embryonic gene expression to study the role of specific proteins in early vertebrate development.

PubMed ID: 10773085
PMC ID: PMC105372
Article link: Nucleic Acids Res
Grant support: [+]

Species referenced: Xenopus
Genes referenced: gs17 pax6

References [+] :
Agrawal, Site-specific excision from RNA by RNase H and mixed-phosphate-backbone oligodeoxynucleotides. 1990, Pubmed