Targeted degradation of mRNA in Xenopus oocytes and embryos directed by modified oligonucleotides: studies of An2 and cyclin in embryogenesis

J M Dagle; J A Walder; D L Weeks

doi:10.1093/nar/18.16.4751

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Targeted degradation of mRNA in Xenopus oocytes and embryos directed by modified oligonucleotides: studies of An2 and cyclin in embryogenesis

Journal article

Open access

Peer reviewed

Targeted degradation of mRNA in Xenopus oocytes and embryos directed by modified oligonucleotides: studies of An2 and cyclin in embryogenesis

J M Dagle, J A Walder and D L Weeks

Nucleic acids research, Vol.18(16), pp.4751-4757

08/25/1990

DOI: 10.1093/nar/18.16.4751

PMCID: PMC331936

PMID: 1697675

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Abstract

We have designed antisense oligodeoxyribonucleotides which are both highly resistant to nucleolytic degradation and also serve as substrates for ribonuclease H. Using these compounds we have targeted the specific degradation of several maternal mRNAs present in Xenopus laevis oocytes and early embryos. Several internucleoside linkages at both the 3' and 5' ends of the oligonucleotides were modified as phosphoramidates to provide complete protection against exonucleases, the predominant nucleolytic activity found in both oocytes and embryos. Eight Internal linkages were left unmodified to provide a substrate for RNase H. Degradation of specific embryonic mRNAs was accomplished using subtoxic amounts of the modified oligonucleotides. Specific depletion of An2, a localized mRNA encoding the alpha subunit of the mitochondrial ATPase, produced embryos that gastrulated later than control embryos and arrested in development prior to neurulation. A modified oligonucleotide targeting Xenopus cyclin B1 and cyclin B2 mRNA was also synthesized. Following the injection of one blastomere of a two-cell embryo with the anti-cyclin oligonucleotide, cell division in that half of the embryo was inhibited, demonstrating the in vivo importance of these cyclins in mitosis. The oligonucleotide analogs described here should be useful in studying developmentally significant proteins in Xenopus.

Mitosis

RNA, Antisense

Mitochondria - enzymology

Oligodeoxyribonucleotides

Endoribonucleases - metabolism

Oocytes - metabolism

RNA, Messenger - genetics

Xenopus laevis

Embryo, Nonmammalian - metabolism

Adenosine Triphosphatases - metabolism

Molecular Sequence Data

Proliferating Cell Nuclear Antigen

Ribonuclease H

RNA, Messenger - metabolism

Animals

Base Sequence

Gastrula - ultrastructure

RNA, Messenger - antagonists & inhibitors

Gastrula - metabolism

Adenosine Triphosphatases - genetics

Exonucleases - metabolism

Nuclear Proteins - genetics

RNA - metabolism

Details

Title: Subtitle: Targeted degradation of mRNA in Xenopus oocytes and embryos directed by modified oligonucleotides: studies of An2 and cyclin in embryogenesis
Creators: J M Dagle - Department of Biochemistry, University of Iowa, Iowa City 52242
J A Walder
D L Weeks
Resource Type: Journal article
Publication Details: Nucleic acids research, Vol.18(16), pp.4751-4757
DOI: 10.1093/nar/18.16.4751
PMID: 1697675
PMCID: PMC331936
NLM abbreviation: Nucleic Acids Res
ISSN: 0305-1048
eISSN: 1362-4962
Publisher: England
Grant note: GM40308 / NIGMS NIH HHS
Language: English
Date published: 08/25/1990
Academic Unit: Stead Family Department of Pediatrics; Epidemiology; Biochemistry and Molecular Biology; Neonatology
Record Identifier: 9984024525502771

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