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PMID: 10468557 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S.

RNA aptamers as effective protein antagonists in a multicellular organism.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 96 ·No. 18 ·1999-08-31 ·Pages 10033-8

Shi H, Hoffman BE, Lis JT

Abstract

RNA aptamers selected against proteins can be used to modulate specific protein function. Expression of such reagents in cells and whole organisms could provide a means of dissecting and controlling molecular mechanisms in vivo. We demonstrate that Drosophila B52 protein can be specifically inhibited in vitro and in vivo by a multivalent RNA aptamer. This inhibitory aptamer RNA binds B52 avidly and inhibits B52-stimulated pre-mRNA splicing. It can be expressed in cultured cells and whole animals in a stable form that accumulates up to 10% of total mRNA. It binds B52 in vivo and suppresses all phenotypes caused by B52 overexpression. The strategies presented here should prove general in design and expression of functional and therapeutic RNAs.

MeSH Terms
Animals Animals, Genetically Modified Base Sequence Cells, Cultured Crosses, Genetic Drosophila/genetics,metabolism Drosophila Proteins Female Genotype Male Molecular Sequence Data Nuclear Proteins/antagonists & inhibitors Nucleic Acid Conformation Oligoribonucleotides/pharmacology Phenotype Phosphoproteins/antagonists & inhibitors RNA/chemistry,genetics RNA Splicing RNA Splicing Factors RNA, Messenger/genetics RNA-Binding Proteins/antagonists & inhibitors Transcription, Genetic
Chemicals
Drosophila Proteins Nuclear Proteins Oligoribonucleotides Phosphoproteins RNA Splicing Factors RNA, Messenger RNA-Binding Proteins B52 protein, Drosophila RNA
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Shi H
Department of Molecular Biology and Genetics, Cornell University, Ithaca, NY 14853, USA.
Hoffman B E
Lis J T
References (39)
39 references, click to expand
  1. The concentration of B52, an essential splicing factor and regulator of splice site choice in vitro, is critical for Drosophila development.
    Mol Cell Biol. 1994 Aug;14(8):5360-70 PMID: 8035814
  2. The superfamily of arginine/serine-rich splicing factors.
    RNA. 1995 Sep;1(7):663-80 PMID: 7585252
  3. Sequence-specific RNA binding by an SR protein requires RS domain phosphorylation: creation of an SRp40-specific splicing enhancer.
    Proc Natl Acad Sci U S A. 1997 Feb 18;94(4):1148-53 PMID: 9037021
  4. Rapid changes in Drosophila transcription after an instantaneous heat shock.
    Mol Cell Biol. 1993 Jun;13(6):3456-63 PMID: 8497261
  5. Targeted gene expression as a means of altering cell fates and generating dominant phenotypes.
    Development. 1993 Jun;118(2):401-15 PMID: 8223268
  6. SR proteins and splicing control.
    Genes Dev. 1996 Jul 1;10(13):1569-79 PMID: 8682289
  7. A structured retroviral RNA element that mediates nucleocytoplasmic export of intron-containing RNA.
    Mol Cell Biol. 1997 Jan;17(1):135-44 PMID: 8972193
  8. Potent and specific genetic interference by double-stranded RNA in Caenorhabditis elegans.
    Nature. 1998 Feb 19;391(6669):806-11 PMID: 9486653
  9. Perturbation of nuclear architecture by long-distance chromosome interactions.
    Cell. 1996 May 31;85(5):745-59 PMID: 8646782
  10. Genetic selection of peptide aptamers that recognize and inhibit cyclin-dependent kinase 2.
    Nature. 1996 Apr 11;380(6574):548-50 PMID: 8606778
  11. Characterization of a Drosophila protein associated with boundaries of transcriptionally active chromatin.
    Genes Dev. 1991 Sep;5(9):1611-21 PMID: 1885003
  12. A good antisense molecule is hard to find.
    Trends Biochem Sci. 1998 Feb;23(2):45-50 PMID: 9538686
  13. Diversity of oligonucleotide functions.
    Annu Rev Biochem. 1995;64:763-97 PMID: 7574500
  14. Systematic evolution of ligands by exponential enrichment: RNA ligands to bacteriophage T4 DNA polymerase.
    Science. 1990 Aug 3;249(4968):505-10 PMID: 2200121
  15. Sex-specific control of Drosophila melanogaster yolk protein 1 gene expression is limited to transcription.
    Mol Cell Biol. 1988 Nov;8(11):4756-64 PMID: 3145403
  16. The human splicing factors ASF/SF2 and SC35 possess distinct, functionally significant RNA binding specificities.
    EMBO J. 1995 Jul 17;14(14):3540-51 PMID: 7543047
  17. Use of dsRNA-mediated genetic interference to demonstrate that frizzled and frizzled 2 act in the wingless pathway.
    Cell. 1998 Dec 23;95(7):1017-26 PMID: 9875855
  18. The white gene as a marker in a new P-element vector for gene transfer in Drosophila.
    Nucleic Acids Res. 1987 May 26;15(10):3947-59 PMID: 3108854
  19. In vivo generation of highly abundant sequence-specific oligonucleotides for antisense and triplex gene regulation.
    Nucleic Acids Res. 1994 Jul 25;22(14):2830-6 PMID: 8052538
  20. Construction of a novel RNA-transcript-trimming plasmid which can be used both in vitro in place of run-off and (G)-free transcriptions and in vivo as multi-sequences transcription vectors.
    Nucleic Acids Res. 1991 Oct 11;19(19):5125-30 PMID: 1923797
  21. Accurate transcription initiation by RNA polymerase II in a soluble extract from isolated mammalian nuclei.
    Nucleic Acids Res. 1983 Mar 11;11(5):1475-89 PMID: 6828386
  22. Selection of efficient cleavage sites in target RNAs by using a ribozyme expression library.
    Mol Cell Biol. 1995 Jan;15(1):540-51 PMID: 7528330
  23. Exceptionally stable nucleic acid hairpins.
    Annu Rev Biophys Biomol Struct. 1995;24:379-404 PMID: 7545040
  24. Targeting cyclin-dependent kinases in Drosophila with peptide aptamers.
    Proc Natl Acad Sci U S A. 1998 Nov 24;95(24):14266-71 PMID: 9826689
  25. Virus resistance and gene silencing in plants can be induced by simultaneous expression of sense and antisense RNA.
    Proc Natl Acad Sci U S A. 1998 Nov 10;95(23):13959-64 PMID: 9811908
  26. Improved accumulation and activity of ribozymes expressed from a tRNA-based RNA polymerase III promoter.
    Nucleic Acids Res. 1995 Jun 25;23(12):2259-68 PMID: 7610054
  27. On finding all suboptimal foldings of an RNA molecule.
    Science. 1989 Apr 7;244(4900):48-52 PMID: 2468181
  28. A consensus sequence polymer inhibits in vivo expression of heat shock genes.
    Mol Cell Biol. 1986 Sep;6(9):3200-6 PMID: 3023967
  29. Theoretical studies of clonal selection: minimal antibody repertoire size and reliability of self-non-self discrimination.
    J Theor Biol. 1979 Dec 21;81(4):645-70 PMID: 94141
  30. Targeted nuclear antisense RNA mimics natural antisense-induced degradation of polyoma virus early RNA.
    Proc Natl Acad Sci U S A. 1994 May 10;91(10):4258-62 PMID: 8183899
  31. The SR protein B52/SRp55 is essential for Drosophila development.
    Mol Cell Biol. 1994 Nov;14(11):7499-506 PMID: 7935465
  32. HSF recruitment and loss at most Drosophila heat shock loci is coordinated and depends on proximal promoter sequences.
    Chromosoma. 1996 Sep;105(3):158-71 PMID: 8781184
  33. Mature mRNA 3' end formation stimulates RNA export from the nucleus.
    EMBO J. 1991 Nov;10(11):3513-22 PMID: 1833188
  34. Varying patterns of protein synthesis in Drosophila during heat shock: implications for regulation.
    Dev Biol. 1980 Jun 15;77(2):463-79 PMID: 7399133
  35. Overexpression of TAR sequences renders cells resistant to human immunodeficiency virus replication.
    Cell. 1990 Nov 2;63(3):601-8 PMID: 2225067
  36. Selective targeting and inhibition of yeast RNA polymerase II by RNA aptamers.
    J Biol Chem. 1997 Oct 31;272(44):27980-6 PMID: 9346949
  37. Simple RNA enzymes with new and highly specific endoribonuclease activities.
    Nature. 1988 Aug 18;334(6183):585-91 PMID: 2457170
  38. Accurate and efficient pre-mRNA splicing in Drosophila cell-free extracts.
    Proc Natl Acad Sci U S A. 1988 May;85(9):2904-8 PMID: 3362856
  39. In vitro selection of RNA molecules that bind specific ligands.
    Nature. 1990 Aug 30;346(6287):818-22 PMID: 1697402
Article Info
Journal
Proceedings of the National Academy of Sciences of the United States of America
Abbr.
Proc Natl Acad Sci U S A
ISSN
0027-8424
DOI
10.1073/pnas.96.18.10033
Published
1999-08-31
Pages
10033-8
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC17837
Subset
IM
Grants
NIGMS NIH HHS · F32 GM017509 · United States
NIGMS NIH HHS · GM17509 · United States
NIGMS NIH HHS · GM40918 · United States
External Links
PubMed source DOI: 10.1073/pnas.96.18.10033 PMC Full Text View journal details
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