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Quantitative hybridization-arrest of mRNA in Xenopus oocytes using single-stranded complementary DNA or oligonucleotide probes.
Kawasaki ES
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The expression of heterologous mRNA in Xenopus oocytes was quantitatively inhibited by coinjection of single-stranded complementary DNA or synthetic complementary oligonucleotides. The lymphokines Interleukin-2 (IL-2) and Interleukin-3 (IL-3) were used as model systems to test the effectiveness of this procedure. Messenger RNA samples were hybridized to single stranded complementary DNA or oligonucleotides, injected into oocytes and the oocyte incubation medium assayed for the presence or absence of specific translation products 48 hours later. When IL-2 mRNA was hybridized to a large excess of long (490 bases) single stranded complementary DNA, the expression of IL-2 was effectively blocked (greater than 98%). Complementary oligonucleotides (18-23 bases) were almost as effective as the polynucleotide in inhibiting IL-2 activity (greater than 95%). Oligonucleotides derived from the 5' end, middle or 3' end of the coding sequence were all effective in arresting IL-2 mRNA translation. Oligonucleotide hybrid-arrest was effective even when no NaCl was present in the hybridization buffer, indicating that the annealing reaction could occur within the oocyte after injection. Definite proof that hybrid-arrest could occur in vivo was shown by the fact that oligonucleotides injected before or after mRNA injection, while not as effective as co-injection, still showed substantial inhibition of specific mRNA translation. The oligonucleotide hybrid-arrest method was equally effective in the case of IL-3, demonstrating its general applicability.
Aviv,
Purification of biologically active globin messenger RNA by chromatography on oligothymidylic acid-cellulose.
1972, Pubmed
Aviv,
Purification of biologically active globin messenger RNA by chromatography on oligothymidylic acid-cellulose.
1972,
Pubmed
Berger,
Inhibition of intractable nucleases with ribonucleoside--vanadyl complexes: isolation of messenger ribonucleic acid from resting lymphocytes.
1979,
Pubmed
Bishop,
Three abundance classes in HeLa cell messenger RNA.
1974,
Pubmed
Chandler,
The use of single-stranded phage DNAs in hybrid arrest and release translation.
1982,
Pubmed
Coleman,
The use of RNAs complementary to specific mRNAs to regulate the expression of individual bacterial genes.
1984,
Pubmed
Devos,
Molecular cloning of human interleukin 2 cDNA and its expression in E. coli.
1983,
Pubmed
Eckhardt,
Blocking of the initiation of protein biosynthesis by a pentanucleotide complementary to the 3' end of Escherichia coli 16 S rRNA.
1979,
Pubmed
Fisher,
Use of complementary DNA oligomers to probe trp leader transcript secondary structures involved in transcription pausing and termination.
1984,
Pubmed
Fung,
Molecular cloning of cDNA for murine interleukin-3.
,
Pubmed
,
Xenbase
Gillis,
T cell growth factor: parameters of production and a quantitative microassay for activity.
1978,
Pubmed
Gurdon,
Use of frog eggs and oocytes for the study of messenger RNA and its translation in living cells.
1971,
Pubmed
,
Xenbase
Harpold,
Construction and identification by positive hybridization-translation of a bacterial plasmid containing a rat growth hormone structural gene sequence.
1978,
Pubmed
Ihle,
Phenotypic characteristics of cell lines requiring interleukin 3 for growth.
1982,
Pubmed
Izant,
Inhibition of thymidine kinase gene expression by anti-sense RNA: a molecular approach to genetic analysis.
1984,
Pubmed
Jayaraman,
Selective inhibition of Escherichia coli protein synthesis and growth by nonionic oligonucleotides complementary to the 3' end of 16S rRNA.
1981,
Pubmed
Lee,
Constitutive production of a unique lymphokine (IL 3) by the WEHI-3 cell line.
1982,
Pubmed
Lerner,
Are snRNPs involved in splicing?
1980,
Pubmed
Mark,
Site-specific mutagenesis of the human fibroblast interferon gene.
1984,
Pubmed
Melton,
Injected anti-sense RNAs specifically block messenger RNA translation in vivo.
1985,
Pubmed
,
Xenbase
Messing,
New M13 vectors for cloning.
1983,
Pubmed
Messing,
A new pair of M13 vectors for selecting either DNA strand of double-digest restriction fragments.
1982,
Pubmed
Mizuno,
A unique mechanism regulating gene expression: translational inhibition by a complementary RNA transcript (micRNA).
1984,
Pubmed
Mount,
The U1 small nuclear RNA-protein complex selectively binds a 5' splice site in vitro.
1983,
Pubmed
Müller,
A homeo-box-containing gene expressed during oogenesis in Xenopus.
1984,
Pubmed
,
Xenbase
Padgett,
Splicing of messenger RNA precursors is inhibited by antisera to small nuclear ribonucleoprotein.
1983,
Pubmed
Paterson,
Structural gene identification and mapping by DNA-mRNA hybrid-arrested cell-free translation.
1977,
Pubmed
Ricciardi,
Purification and mapping of specific mRNAs by hybridization-selection and cell-free translation.
1979,
Pubmed
Rogers,
A mechanism for RNA splicing.
1980,
Pubmed
Rosenberg,
Production of phenocopies by Krüppel antisense RNA injection into Drosophila embryos.
,
Pubmed
Rosenberg,
Biological activity of recombinant human interleukin-2 produced in Escherichia coli.
1984,
Pubmed
Simons,
Translational control of IS10 transposition.
1983,
Pubmed
Stephenson,
Inhibition of Rous sarcoma viral RNA translation by a specific oligodeoxyribonucleotide.
1978,
Pubmed
Taniguchi,
Inhibition of Qbeta RNA 70S ribosome initiation complex formation by an oligonucleotide complementary to the 3' terminal region of E. coli 16S ribosomal RNA.
1978,
Pubmed
Taniguchi,
Structure and expression of a cloned cDNA for human interleukin-2.
,
Pubmed
Tomizawa,
Plasmid ColE1 incompatibility determined by interaction of RNA I with primer transcript.
1981,
Pubmed
Wang,
Site-specific mutagenesis of the human interleukin-2 gene: structure-function analysis of the cysteine residues.
1984,
Pubmed
Winkler,
Transcription termination at the tryptophan operon attenuator is decreased in vitro by an oligomer complementary to a segment of the leader transcript.
1982,
Pubmed
Yokota,
Isolation and characterization of a mouse cDNA clone that expresses mast-cell growth-factor activity in monkey cells.
1984,
Pubmed
,
Xenbase
Zamecnik,
Inhibition of Rous sarcoma virus replication and cell transformation by a specific oligodeoxynucleotide.
1978,
Pubmed
