Home LiteratureArticle Details
PMID: 11481446 Published · ppublish English Comparative Study Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Specific inhibition of gene expression by small double-stranded RNAs in invertebrate and vertebrate systems.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 98 ·No. 17 ·2001-08-14 ·Pages 9742-7

Caplen NJ, Parrish S, Imani F, Fire A, Morgan RA

Abstract

Short interfering RNAs (siRNAs) are double-stranded RNAs of approximately 21-25 nucleotides that have been shown to function as key intermediaries in triggering sequence-specific RNA degradation during posttranscriptional gene silencing in plants and RNA interference in invertebrates. siRNAs have a characteristic structure, with 5'-phosphate/3'-hydroxyl ends and a 2-base 3' overhang on each strand of the duplex. In this study, we present data that synthetic siRNAs can induce gene-specific inhibition of expression in Caenorhabditis elegans and in cell lines from humans and mice. In each case, the interference by siRNAs was superior to the inhibition of gene expression mediated by single-stranded antisense oligonucleotides. The siRNAs seem to avoid the well documented nonspecific effects triggered by longer double-stranded RNAs in mammalian cells. These observations may open a path toward the use of siRNAs as a reverse genetic and therapeutic tool in mammalian cells.

MeSH Terms
Animals Caenorhabditis elegans/genetics,metabolism Caenorhabditis elegans Proteins Calmodulin-Binding Proteins/biosynthesis,genetics Cell Death Cells, Cultured/drug effects,metabolism Chloramphenicol O-Acetyltransferase/biosynthesis,genetics Endoribonucleases/metabolism Gene Silencing/drug effects,physiology Genes, Reporter Green Fluorescent Proteins HeLa Cells/drug effects,metabolism Helminth Proteins/biosynthesis,genetics Humans Invertebrates/genetics Luminescent Proteins/biosynthesis,genetics Mammals/genetics Mice Muscle Proteins/biosynthesis,genetics Phosphorylation RNA, Antisense/chemical synthesis,pharmacology RNA, Double-Stranded/pharmacology,physiology RNA, Helminth/physiology Recombinant Fusion Proteins/biosynthesis Ribonuclease III Species Specificity Transfection Vertebrates/genetics
Chemicals
Caenorhabditis elegans Proteins Calmodulin-Binding Proteins Helminth Proteins Luminescent Proteins Muscle Proteins RNA, Antisense RNA, Double-Stranded RNA, Helminth Recombinant Fusion Proteins unc-22 protein, C elegans Green Fluorescent Proteins Chloramphenicol O-Acetyltransferase Endoribonucleases Ribonuclease III
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Caplen N J
Medical Genetics Branch, National Human Genome Research Institute, National Institutes of Health, Bethesda, MD 20892, USA. [email protected]
Parrish S
Imani F
Fire A
Morgan R A
References (50)
50 references, click to expand
  1. Double-stranded RNA as a template for gene silencing.
    Cell. 2000 Apr 28;101(3):235-8 PMID: 10847677
  2. EGO-1 is related to RNA-directed RNA polymerase and functions in germ-line development and RNA interference in C. elegans.
    Curr Biol. 2000 Feb 24;10(4):169-78 PMID: 10704412
  3. An RNA-dependent RNA polymerase gene in Arabidopsis is required for posttranscriptional gene silencing mediated by a transgene but not by a virus.
    Cell. 2000 May 26;101(5):543-53 PMID: 10850496
  4. The evolutionarily conserved BMP-binding protein Twisted gastrulation promotes BMP signalling.
    Nature. 2000 Jun 15;405(6788):757-63 PMID: 10866189
  5. Transgene-mediated cosuppression in the C. elegans germ line.
    Genes Dev. 2000 Jul 1;14(13):1578-83 PMID: 10887151
  6. Too much interference: injection of double-stranded RNA has nonspecific effects in the zebrafish embryo.
    Dev Biol. 2000 Aug 1;224(1):20-8 PMID: 10898958
  7. RNAi--prospects for a general technique for determining gene function.
    Parasitol Today. 2000 Aug;16(8):347-9 PMID: 10900483
  8. dsRNA-mediated gene silencing in cultured Drosophila cells: a tissue culture model for the analysis of RNA interference.
    Gene. 2000 Jul 11;252(1-2):95-105 PMID: 10903441
  9. RNA interference for the organizer-specific gene Xlim-1 in Xenopus embryos.
    Biochem Biophys Res Commun. 2000 Aug 2;274(2):434-9 PMID: 10913356
  10. Selective reduction of dormant maternal mRNAs in mouse oocytes by RNA interference.
    Development. 2000 Oct;127(19):4147-56 PMID: 10976047
  11. Sensitive assay of RNA interference in Drosophila and Chinese hamster cultured cells using firefly luciferase gene as target.
    FEBS Lett. 2000 Aug 18;479(3):79-82 PMID: 10981711
  12. A link between RNA interference and nonsense-mediated decay in Caenorhabditis elegans.
    Science. 2000 Sep 15;289(5486):1928-31 PMID: 10988072
  13. Evidence that processed small dsRNAs may mediate sequence-specific mRNA degradation during RNAi in Drosophila embryos.
    Curr Biol. 2000 Oct 5;10(19):1191-200 PMID: 11050387
  14. Transgene and transposon silencing in Chlamydomonas reinhardtii by a DEAH-box RNA helicase.
    Science. 2000 Nov 10;290(5494):1159-62 PMID: 11073454
  15. Human RNase III is a 160-kDa protein involved in preribosomal RNA processing.
    J Biol Chem. 2000 Nov 24;275(47):36957-65 PMID: 10948199
  16. Post-transcriptional gene silencing across kingdoms.
    Curr Opin Genet Dev. 2000 Dec;10(6):638-43 PMID: 11088014
  17. Virus-encoded suppressor of posttranscriptional gene silencing targets a maintenance step in the silencing pathway.
    Proc Natl Acad Sci U S A. 2000 Nov 21;97(24):13401-6 PMID: 11078509
  18. Double-stranded RNA injection produces nonspecific defects in zebrafish.
    Dev Biol. 2001 Jan 1;229(1):215-23 PMID: 11133165
  19. Role for a bidentate ribonuclease in the initiation step of RNA interference.
    Nature. 2001 Jan 18;409(6818):363-6 PMID: 11201747
  20. RNA interference is mediated by 21- and 22-nucleotide RNAs.
    Genes Dev. 2001 Jan 15;15(2):188-200 PMID: 11157775
  21. RNA interference--2001.
    Genes Dev. 2001 Mar 1;15(5):485-90 PMID: 11238371
  22. Gene silencing by double-stranded RNA.
    Curr Opin Cell Biol. 2001 Apr;13(2):244-8 PMID: 11248560
  23. RNA-based silencing strategies in plants.
    Curr Opin Genet Dev. 2001 Apr;11(2):221-7 PMID: 11250148
  24. Duplexes of 21-nucleotide RNAs mediate RNA interference in cultured mammalian cells.
    Nature. 2001 May 24;411(6836):494-8 PMID: 11373684
  25. Functional anatomy of a dsRNA trigger: differential requirement for the two trigger strands in RNA interference.
    Mol Cell. 2000 Nov;6(5):1077-87 PMID: 11106747
  26. Characteristics of a human cell line transformed by DNA from human adenovirus type 5.
    J Gen Virol. 1977 Jul;36(1):59-74 PMID: 886304
  27. Structural requirements of double-stranded RNA for the activation of 2',5'-oligo(A) polymerase and protein kinase of interferon-treated HeLa cells.
    J Biol Chem. 1979 Oct 25;254(20):10180-3 PMID: 489592
  28. Interactions between double-stranded RNA regulators and the protein kinase DAI.
    Mol Cell Biol. 1992 Nov;12(11):5238-48 PMID: 1357546
  29. The double-stranded RNA-dependent protein kinase PKR: structure and function.
    J Interferon Cytokine Res. 1997 Sep;17(9):503-24 PMID: 9335428
  30. The 2-5A system: modulation of viral and cellular processes through acceleration of RNA degradation.
    Pharmacol Ther. 1998 May;78(2):55-113 PMID: 9623881
  31. A counterdefensive strategy of plant viruses: suppression of posttranscriptional gene silencing.
    Cell. 1998 Nov 13;95(4):461-70 PMID: 9827799
  32. Characterization of transgenic mice with targeted disruption of the catalytic domain of the double-stranded RNA-dependent protein kinase, PKR.
    J Biol Chem. 1999 Feb 26;274(9):5953-62 PMID: 10026221
  33. Gene silencing in Neurospora crassa requires a protein homologous to RNA-dependent RNA polymerase.
    Nature. 1999 May 13;399(6732):166-9 PMID: 10335848
  34. RNA-triggered gene silencing.
    Trends Genet. 1999 Sep;15(9):358-63 PMID: 10461204
  35. Nonsense-mediated mRNA decay in health and disease.
    Hum Mol Genet. 1999;8(10):1893-900 PMID: 10469842
  36. Double-stranded RNA induces specific developmental defects in zebrafish embryos.
    Biochem Biophys Res Commun. 1999 Sep 16;263(1):156-61 PMID: 10486270
  37. Epigenetics: regulation through repression.
    Science. 1999 Oct 15;286(5439):481-6 PMID: 10521337
  38. Quantitative studies of the growth of mouse embryo cells in culture and their development into established lines.
    J Cell Biol. 1963 May;17:299-313 PMID: 13985244
  39. The rde-1 gene, RNA interference, and transposon silencing in C. elegans.
    Cell. 1999 Oct 15;99(2):123-32 PMID: 10535731
  40. Mut-7 of C. elegans, required for transposon silencing and RNA interference, is a homolog of Werner syndrome helicase and RNaseD.
    Cell. 1999 Oct 15;99(2):133-41 PMID: 10535732
  41. A species of small antisense RNA in posttranscriptional gene silencing in plants.
    Science. 1999 Oct 29;286(5441):950-2 PMID: 10542148
  42. An RNA-directed nuclease mediates post-transcriptional gene silencing in Drosophila cells.
    Nature. 2000 Mar 16;404(6775):293-6 PMID: 10749213
  43. mRNA stability in eukaryotes.
    Curr Opin Genet Dev. 2000 Apr;10(2):193-8 PMID: 10753781
  44. RNAi: double-stranded RNA directs the ATP-dependent cleavage of mRNA at 21 to 23 nucleotide intervals.
    Cell. 2000 Mar 31;101(1):25-33 PMID: 10778853
  45. Molecular cloning and characterization of a novel human gene (HERNA) which encodes a putative RNA-helicase.
    Biochim Biophys Acta. 2000 Jan 31;1490(1-2):163-9 PMID: 10786632
  46. Molecular mechanisms of action of antisense drugs.
    Biochim Biophys Acta. 1999 Dec 10;1489(1):31-44 PMID: 10806995
  47. Targeted mRNA degradation by double-stranded RNA in vitro.
    Genes Dev. 1999 Dec 15;13(24):3191-7 PMID: 10617568
  48. Double-stranded RNA injection produces null phenotypes in zebrafish.
    Dev Biol. 2000 Jan 15;217(2):394-405 PMID: 10625563
  49. Specific interference with gene function by double-stranded RNA in early mouse development.
    Nat Cell Biol. 2000 Feb;2(2):70-5 PMID: 10655585
  50. Arabidopsis SGS2 and SGS3 genes are required for posttranscriptional gene silencing and natural virus resistance.
    Cell. 2000 May 26;101(5):533-42 PMID: 10850495
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.171251798
Email
[email protected]
Published
2001-08-14
Epub
2001-00-31
Pages
9742-7
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC55523
Subset
IM
Grants
NIGMS NIH HHS · R01 GM037706 · United States
NIGMS NIH HHS · AF R01-GM37706 · United States
NIAID NIH HHS · FI AI446962 · United States
NIGMS NIH HHS · T32-GM07321 · United States
External Links
PubMed source DOI: 10.1073/pnas.171251798 PMC Full Text View journal details
Analysis Services
Analysis Services

Contact

No. 2 Wenbo Road, Zhangqiu District, Jinan, Shandong

Qilu Normal University · Genelibs Bioinformatics Lab

750 Shunhua Rd, Jinan

2F, Bldg F, University Science Park

Tel: 0531-88819269

WeChat Official Account

Follow our WeChat subscription account for real-time updates and the latest in medical and biological research.


Business Email

E-mail: [email protected]

Copyright © 2026.Genelibs Jinan Honghui Biotechnology Co., Ltd. 鲁ICP备20007159号-2