RNA Interference: Its Use as Antiviral Therapy

RNA interference (RNAi) is a sequence-specific gene-silencing mechanism that has been proposed to function as a defence mechanism of eukaryotic cells against viruses and transposons. RNAi was first observed in plants in the form of a mysterious immune response to viral pathogens. But RNAi is more than just a response to exogenous genetic material. Small RNAs termed microRNA (miRNA) regulate cellular gene expression programs to control diverse steps in cell development and physiology. The discovery that exogenously delivered short interfering RNA (siRNA) can trigger RNAi in mammalian cells has made it into a powerful technique for generating genetic knock-outs. It also raises the possibility to use RNAi technology as a therapeutic tool against pathogenic viruses. Indeed, inhibition of virus replication has been reported for several human pathogens including human immunodeficiency virus, the hepatitis B and C viruses and influenza virus. We reviewed the field of antiviral RNAi research in 2003 (Haasnoot et al. 2003), but many new studies have recently been published. In this review, we present a complete listing of all antiviral strategies published up to and including December 2004. The latest developments in the RNAi field and their antiviral application are described.

[1]  B. Ramratnam,et al.  Promoter choice affects the potency of HIV-1 specific RNA interference. , 2003, Nucleic acids research.

[2]  B. Ramratnam,et al.  Nucleotide sequence homology requirements of HIV-1-specific short hairpin RNA. , 2003, Nucleic acids research.

[3]  Anindya Dutta,et al.  Small RNAs with Imperfect Match to Endogenous mRNA Repress Translation , 2003, Journal of Biological Chemistry.

[4]  Anton P. McCaffrey,et al.  Inhibition of hepatitis B virus in mice by RNA interference , 2003, Nature Biotechnology.

[5]  M. Mathews,et al.  Interactions between double-stranded RNA regulators and the protein kinase DAI , 1992, Molecular and cellular biology.

[6]  Xiaodong Wang,et al.  R2D2, a Bridge Between the Initiation and Effector Steps of the Drosophila RNAi Pathway , 2003, Science.

[7]  Ben Berkhout,et al.  HIV-1 can escape from RNA interference by evolving an alternative structure in its RNA genome , 2005, Nucleic acids research.

[8]  Brenda L Bass,et al.  Mutations in RNAi Rescue Aberrant Chemotaxis of ADAR Mutants , 2003, Science.

[9]  Michael T. McManus,et al.  RNA interference of influenza virus production by directly targeting mRNA for degradation and indirectly inhibiting all viral RNA transcription , 2003, Proceedings of the National Academy of Sciences of the United States of America.

[10]  B. Clotet,et al.  Suppression of chemokine receptor expression by RNA interference allows for inhibition of HIV-1 replication , 2002, AIDS.

[11]  J. M. Thomson,et al.  Argonaute2 Is the Catalytic Engine of Mammalian RNAi , 2004, Science.

[12]  Zhi Chen,et al.  Inhibition of human La protein by RNA interference downregulates hepatitis B virus mRNA in 2.2.15 cells. , 2004, World journal of gastroenterology.

[13]  S. Barik,et al.  Phenotypic silencing of cytoplasmic genes using sequence-specific double-stranded short interfering RNA and its application in the reverse genetics of wild type negative-strand RNA viruses , 2001, BMC Microbiology.

[14]  K. Taira,et al.  Control of siRNA expression using the Cre-loxP recombination system. , 2004, Nucleic acids research.

[15]  K. Saksela.  Human viruses under attack by small inhibitory RNA. , 2003, Trends in microbiology.

[16]  B. Ramratnam,et al.  Enhanced gene silencing of HIV-1 specific siRNA using microRNA designed hairpins. , 2004, Nucleic acids research.

[17]  T. Du,et al.  Asymmetry in the Assembly of the RNAi Enzyme Complex , 2003, Cell.

[18]  Harry Smith,et al.  Phytochromes and light signal perception by plants—an emerging synthesis , 2000, Nature.

[19]  M. Katze,et al.  Molecular mechanisms of interferon resistance mediated by viral-directed inhibition of PKR, the interferon-induced protein kinase. , 1998, Pharmacology & therapeutics.

[20]  Wei-zao Chen,et al.  RNA Interference Targeting VP1 Inhibits Foot-and-Mouth Disease Virus Replication in BHK-21 Cells and Suckling Mice , 2004, Journal of Virology.

[21]  P. Sharp,et al.  Embryonic stem cell-specific MicroRNAs. , 2003, Developmental cell.

[22]  T. Rana,et al.  Inhibition of Human Immunodeficiency Virus Type 1 Replication by RNA Interference Directed against Human Transcription Elongation Factor P-TEFb (CDK9/CyclinT1) , 2004, Journal of Virology.

[23]  T. Tumpey,et al.  Protection against lethal influenza virus challenge by RNA interference in vivo. , 2004, Proceedings of the National Academy of Sciences of the United States of America.

[24]  Neil A. Smith,et al.  Gene expression: Total silencing by intron-spliced hairpin RNAs , 2000, Nature.

[25]  T. Du,et al.  RISC Assembly Defects in the Drosophila RNAi Mutant armitage , 2004, Cell.

[26]  H. Vaucheret,et al.  RNA Silencing in Plants--Defense and Counterdefense , 2001, Science.

[27]  C. Samuel,et al.  Adenovirus VAI RNA antagonizes the antiviral action of interferon by preventing activation of the interferon-induced eIF-2α kinase , 1986, Cell.

[28]  D. Weissman,et al.  Inhibition of HIV-1 Infection by Small Interfering RNA-Mediated RNA Interference1 , 2002, The Journal of Immunology.

[29]  T. Decker,et al.  Phosphorylation of the Stat1 transactivating domain is required for the response to type I interferons , 2003, EMBO reports.

[30]  M. Oshimura,et al.  Dicer is essential for formation of the heterochromatin structure in vertebrate cells , 2004, Nature Cell Biology.

[31]  Boro Dropulic,et al.  Antisense-Mediated Inhibition of Human Immunodeficiency Virus (HIV) Replication by Use of an HIV Type 1-Based Vector Results in Severely Attenuated Mutants Incapable of Developing Resistance , 2004, Journal of Virology.

[32]  E. Moss,et al.  Conservation of the heterochronic regulator Lin-28, its developmental expression and microRNA complementary sites. , 2003, Developmental biology.

[33]  B. Berkhout,et al.  Inhibition of virus replication by RNA interference. , 2003, Journal of biomedical science.

[34]  Z. Adelman,et al.  RNA Silencing of Dengue Virus Type 2 Replication in Transformed C6/36 Mosquito Cells Transcribing an Inverted-Repeat RNA Derived from the Virus Genome , 2002, Journal of Virology.

[35]  S. Elledge,et al.  Dicer is essential for mouse development , 2003, Nature Genetics.

[36]  B. Cullen,et al.  Human microRNAs are processed from capped, polyadenylated transcripts that can also function as mRNAs. , 2004, RNA.

[37]  C. Mello,et al.  The dsRNA Binding Protein RDE-4 Interacts with RDE-1, DCR-1, and a DExH-Box Helicase to Direct RNAi in C. elegans , 2002, Cell.

[38]  D. Bartel.  MicroRNAs Genomics, Biogenesis, Mechanism, and Function , 2004, Cell.

[39]  T. Tuschl,et al.  Identification of Tissue-Specific MicroRNAs from Mouse , 2002, Current Biology.

[40]  Yutaka Tsutsumi,et al.  HIV-1 nef suppression by virally encoded microRNA , 2004, Retrovirology.

[41]  F. Bushman,et al.  Inhibition of Retroviral Pathogenesis by RNA Interference , 2002, Current Biology.

[42]  N. Caplen,et al.  Inhibition of viral gene expression and replication in mosquito cells by dsRNA-triggered RNA interference. , 2002, Molecular therapy : the journal of the American Society of Gene Therapy.

[43]  Y. Guan,et al.  Inhibition of SARS-associated coronavirus infection and replication by RNA interference. , 2003, JAMA.

[44]  F. Holstege,et al.  Specific inhibition of gene expression using a stably integrated, inducible small‐interfering‐RNA vector , 2003, EMBO reports.

[45]  B. Williams,et al.  PKR; a sentinel kinase for cellular stress , 1999, Oncogene.

[46]  Jiadong Wang,et al.  siRNA-mediated inhibition of HBV replication and expression. , 2004, World journal of gastroenterology.

[47]  B. Cullen,et al.  Potent and Specific Inhibition of Human Immunodeficiency Virus Type 1 Replication by RNA Interference , 2002, Journal of Virology.

[48]  A. Fire,et al.  Potent and specific genetic interference by double-stranded RNA in Caenorhabditis elegans , 1998, Nature.

[49]  J. Lieberman,et al.  Sustained Small Interfering RNA-Mediated HumanImmunodeficiency Virus Type 1 Inhibition in PrimaryMacrophages , 2003, Journal of Virology.

[50]  M. Mathews,et al.  A new species of virus-coded low molecular weight RNA from cells infected with adenovirus type 2 , 1976, Cell.

[51]  Eun-Young Choi,et al.  The C. elegans microRNA let-7 binds to imperfect let-7 complementary sites from the lin-41 3'UTR. , 2004, Genes & development.

[52]  A. Fire,et al.  Specific inhibition of gene expression by small double-stranded RNAs in invertebrate and vertebrate systems , 2001, Proceedings of the National Academy of Sciences of the United States of America.

[53]  Henning Urlaub,et al.  Single-Stranded Antisense siRNAs Guide Target RNA Cleavage in RNAi , 2002, Cell.

[54]  A. Pasquinelli,et al.  Genes and Mechanisms Related to RNA Interference Regulate Expression of the Small Temporal RNAs that Control C. elegans Developmental Timing , 2001, Cell.

[55]  B. Berkhout.  RNA interference as an antiviral approach: targeting HIV-1. , 2004, Current opinion in molecular therapeutics.

[56]  Hajime Sakai,et al.  Regulation of Flowering Time and Floral Organ Identity by a MicroRNA and Its APETALA2-Like Target Genes Article, publication date, and citation information can be found at www.plantcell.org/cgi/doi/10.1105/tpc.016238. , 2003, The Plant Cell Online.

[57]  B. Cullen,et al.  Inhibition of Human Immunodeficiency Virus Type 1 Replication in Primary Macrophages by Using Tat- or CCR5-Specific Small Interfering RNAs Expressed from a Lentivirus Vector , 2003, Journal of Virology.

[58]  T. Tuschl,et al.  RNA interference is mediated by 21- and 22-nucleotide RNAs. , 2001, Genes & development.

[59]  P. Waterhouse,et al.  Virus resistance and gene silencing in plants can be induced by simultaneous expression of sense and antisense RNA. , 1998, Proceedings of the National Academy of Sciences of the United States of America.

[60]  Anastasia Khvorova,et al.  RNA interference blocks gene expression and RNA synthesis from hepatitis C replicons propagated in human liver cells , 2003, Proceedings of the National Academy of Sciences of the United States of America.

[61]  Lin Deng,et al.  Suppression of Hepatitis C Virus Replicon by RNA Interference Directed against the NS3 and NS5B Regions of the Viral Genome , 2004, Microbiology and immunology.

[62]  L. Babiuk,et al.  Silencing SARS‐CoV Spike protein expression in cultured cells by RNA interference , 2004, FEBS Letters.

[63]  G. Hutvagner,et al.  A microRNA in a Multiple-Turnover RNAi Enzyme Complex , 2002, Science.

[64]  Y. Shaul,et al.  Inhibition of hepatitis B virus expression and replication by RNA interference , 2003, Hepatology.

[65]  J. Rossi,et al.  Inhibition of HIV-1 infection by lentiviral vectors expressing Pol III-promoted anti-HIV RNAs. , 2003, Molecular therapy : the journal of the American Society of Gene Therapy.

[66]  J. Rossi,et al.  Negative feedback inhibition of HIV-1 by TAT-inducible expression of siRNA , 2004, Nature Biotechnology.

[67]  B. Ramratnam,et al.  Human Immunodeficiency Virus Type 1 Escape from RNA Interference , 2003, Journal of Virology.

[68]  B. Berkhout,et al.  Identification of a new human coronavirus , 2004, Nature Medicine.

[69]  R. Pomerantz,et al.  Antiviral Effects of Human Immunodeficiency Virus Type 1-Specific Small Interfering RNAs against Targets Conserved in Select Neurotropic Viral Strains , 2004, Journal of Virology.

[70]  K. Eguchi,et al.  Short interfering RNA‐directed inhibition of hepatitis B virus replication , 2003, FEBS letters.

[71]  D. Baltimore,et al.  Inhibiting HIV-1 infection in human T cells by lentiviral-mediated delivery of small interfering RNA against CCR5 , 2002, Proceedings of the National Academy of Sciences of the United States of America.

[72]  Ben Berkhout,et al.  Human Immunodeficiency Virus Type 1 Escapes from RNA Interference-Mediated Inhibition , 2004, Journal of Virology.

[73]  Kuan-Teh Jeang,et al.  HIV-1 encoded candidate micro-RNAs and their cellular targets , 2004, Retrovirology.

[74]  E. Moss,et al.  Small-interfering RNAs in the radar of the interferon system , 2003, Nature Cell Biology.

[75]  J. Rossi,et al.  Rapid assessment of anti-HIV siRNA efficacy using PCR-derived Pol III shRNA cassettes. , 2004, Molecular therapy : the journal of the American Society of Gene Therapy.

[76]  Hiroyuki Miyoshi,et al.  Optimization of an siRNA‐expression system with an improved hairpin and its significant suppressive effects in mammalian cells , 2004, The journal of gene medicine.

[77]  B. Cullen,et al.  Exportin-5 mediates the nuclear export of pre-microRNAs and short hairpin RNAs. , 2003, Genes & development.

[78]  H. Iwata,et al.  Inhibition of Virus Production in JC Virus-Infected Cells by Postinfection RNA Interference , 2004, Journal of Virology.

[79]  B. Li,et al.  Expression profiling reveals off-target gene regulation by RNAi , 2003, Nature Biotechnology.

[80]  L. Ren,et al.  Inhibition of Severe Acute Respiratory Syndrome Virus Replication by Small Interfering RNAs in Mammalian Cells , 2004, Journal of Virology.

[81]  Edwin Cuppen,et al.  The microRNA-producing enzyme Dicer1 is essential for zebrafish development , 2003, Nature Genetics.

[82]  N. Saksena,et al.  Double‐Stranded nef RNA Interferes with Human Immunodeficiency Virus Type 1 Replication , 2002, Microbiology and immunology.

[83]  N. Miyano-Kurosaki,et al.  Prevention of HIV-1 infection in human peripheral blood mononuclear cells by specific RNA interference. , 2002, Nucleic acids research.

[84]  Y. Li,et al.  Genome-wide expression profiling of RNA interference of hepatitis B virus gene expression and replication , 2004, Cellular and Molecular Life Sciences CMLS.

[85]  S. Kung,et al.  Selective inhibition of enterovirus 71 replication by short hairpin RNAs. , 2004, Biochemical and biophysical research communications.

[86]  R. Ray,et al.  Inhibition of hepatitis C virus protein expression by RNA interference. , 2003, Virus research.

[87]  Qing Ge,et al.  Inhibition of influenza virus production in virus-infected mice by RNA interference. , 2004, Proceedings of the National Academy of Sciences of the United States of America.

[88]  T. Rana,et al.  Modulating HIV-1 replication by RNA interference directed against human transcription elongation factor SPT5 , 2004, Retrovirology.

[89]  Nick V Grishin,et al.  Biochemical identification of Argonaute 2 as the sole protein required for RNA-induced silencing complex activity. , 2004, Proceedings of the National Academy of Sciences of the United States of America.

[90]  R. Bernards,et al.  A System for Stable Expression of Short Interfering RNAs in Mammalian Cells , 2002, Science.

[91]  V. Ambros,et al.  An Extensive Class of Small RNAs in Caenorhabditis elegans , 2001, Science.

[92]  C. Arias,et al.  Rotavirus gene silencing by small interfering RNAs , 2002, EMBO reports.

[93]  D. Headon,et al.  A Link Between mRNA Turnover and RNA Interference in Arabidopsis , 2004, Science.

[94]  Jinhong Chang,et al.  Susceptibility of Human Hepatitis Delta Virus RNAs to Small Interfering RNA Action , 2003, Journal of Virology.

[95]  A. Pasquinelli,et al.  A Cellular Function for the RNA-Interference Enzyme Dicer in the Maturation of the let-7 Small Temporal RNA , 2001, Science.

[96]  Edwards Allen,et al.  P1/HC-Pro, a viral suppressor of RNA silencing, interferes with Arabidopsis development and miRNA unction. , 2003, Developmental cell.

[97]  R. Surabhi,et al.  RNA Interference Directed against Viral and Cellular Targets Inhibits Human Immunodeficiency Virus Type 1 Replication , 2002, Journal of Virology.

[98]  Hong Yang,et al.  Inhibition of respiratory syncytial virus infection with intranasal siRNA nanoparticles targeting the viral NS1 gene , 2005, Nature Medicine.

[99]  M. Mann,et al.  miRNPs: a novel class of ribonucleoproteins containing numerous microRNAs. , 2002, Genes & development.

[100]  Ali Ehsani,et al.  Expression of small interfering RNAs targeted against HIV-1 rev transcripts in human cells , 2002, Nature Biotechnology.

[101]  K. Khalili,et al.  Intracellular Approach for Blocking JC Virus Gene Expression by Using RNA Interference during Viral Infection , 2004, Journal of Virology.

[102]  A. Banerjea,et al.  Bispecific short hairpin siRNA constructs targeted to CD4, CXCR4, and CCR5 confer HIV-1 resistance. , 2003, Oligonucleotides.

[103]  Ben Berkhout,et al.  Suppression of RNA Interference by Adenovirus Virus-Associated RNA , 2005, Journal of Virology.

[104]  P. Bevilacqua,et al.  Straightening of bulged RNA by the double-stranded RNA-binding domain from the protein kinase PKR. , 2000, Proceedings of the National Academy of Sciences of the United States of America.

[105]  M. Strand,et al.  RNA interference silences Microplitis demolitor bracovirus genes and implicates glc1.8 in disruption of adhesion in infected host cells. , 2003, Virology.

[106]  J. Milner,et al.  Selective silencing of viral gene expression in HPV-positive human cervical carcinoma cells treated with siRNA, a primer of RNA interference , 2002, Oncogene.

[107]  S. Fesik,et al.  Specificity of short interfering RNA determined through gene expression signatures , 2003, Proceedings of the National Academy of Sciences of the United States of America.

[108]  T. Klimkait,et al.  Inhibition of HIV-1 multiplication by antisense U7 snRNAs and siRNAs targeting cyclophilin A. , 2004, Nucleic acids research.

[109]  P. Green,et al.  AtXRN4 degrades mRNA in Arabidopsis and its substrates include selected miRNA targets. , 2004, Molecular cell.

[110]  Tyra G. Wolfsberg,et al.  Short interfering RNAs can induce unexpected and divergent changes in the levels of untargeted proteins in mammalian cells , 2004, Proceedings of the National Academy of Sciences of the United States of America.

[111]  Phillip D. Zamore,et al.  RNA Interference , 2000, Science.

[112]  G. Hannon,et al.  C . elegans involved in developmental timing in Dicer functions in RNA interference and in synthesis of small RNA , 2001 .

[113]  M. Mathews.  Structure, function, and evolution of adenovirus virus-associated RNAs. , 1995, Current topics in microbiology and immunology.

[114]  G. Akusjärvi,et al.  Adenovirus VA RNAI: a positive regulator of mRNA translation , 1984, Molecular and cellular biology.

[115]  R. Sun,et al.  Inhibition of Gammaherpesvirus Replication by RNA Interference , 2003, Journal of Virology.

[116]  Matthias John,et al.  Therapeutic silencing of an endogenous gene by systemic administration of modified siRNAs , 2004, Nature.

[117]  M. Wiznerowicz,et al.  Conditional Suppression of Cellular Genes: Lentivirus Vector-Mediated Drug-Inducible RNA Interference , 2003, Journal of Virology.

[118]  G. Hannon.  RNA interference : RNA , 2002 .

[119]  Konstantin Khrapko,et al.  A microRNA array reveals extensive regulation of microRNAs during brain development. , 2003, RNA.

[120]  C. Arias,et al.  Silencing the Morphogenesis of Rotavirus , 2005, Journal of Virology.

[121]  R. Andino,et al.  Short interfering RNA confers intracellular antiviral immunity in human cells , 2002, Nature.

[122]  Mamoru Watanabe,et al.  Inhibition of intracellular hepatitis C virus replication by synthetic and vector‐derived small interfering RNAs , 2003, EMBO reports.

[123]  Pasko Rakic,et al.  Microarray analysis of microRNA expression in the developing mammalian brain , 2004, Genome Biology.

[124]  Akira Ishizuka,et al.  Distinct roles for Argonaute proteins in small RNA-directed RNA cleavage pathways. , 2004, Genes & development.

[125]  Robert H. Silverman,et al.  Activation of the interferon system by short-interfering RNAs , 2003, Nature Cell Biology.

[126]  F. Gannon.  Educate or communicate? | Ring out the old, ring in the new , 2003 .

[127]  R. Iggo,et al.  Induction of an interferon response by RNAi vectors in mammalian cells , 2003, Nature Genetics.

[128]  B. Samuelsson,et al.  Ribonuclease activity and RNA binding of recombinant human Dicer , 2002, The EMBO journal.

[129]  R. J. Clem,et al.  Silencing of the Baculovirus Op-iap3 Gene by RNA Interference Reveals that It Is Required for Prevention of Apoptosis during Orgyia pseudotsugata M Nucleopolyhedrovirus Infection of Ld652Y Cells , 2003, Journal of Virology.

[130]  Shih-Shin Chang,et al.  Inhibition of the Epstein-Barr virus lytic cycle by Zta-targeted RNA interference. , 2004, The Journal of general virology.

[131]  B. Ramratnam,et al.  Efficient gene transfer of HIV-1-specific short hairpin RNA into human lymphocytic cells using recombinant adeno-associated virus vectors. , 2004, Molecular therapy : the journal of the American Society of Gene Therapy.

[132]  B. Schaffhausen,et al.  Attenuation of SARS coronavirus by a short hairpin RNA expression plasmid targeting RNA-dependent RNA polymerase , 2004, Virology.

[133]  D. Engelke,et al.  Effective expression of small interfering RNA in human cells , 2002, Nature Biotechnology.

[134]  W. Filipowicz,et al.  Tethering of human Ago proteins to mRNA mimics the miRNA-mediated repression of protein synthesis. , 2004, RNA.

[135]  Michael R. Green,et al.  Nonspecific, concentration-dependent stimulation and repression of mammalian gene expression by small interfering RNAs (siRNAs). , 2004, RNA.

[136]  Anton J. Enright,et al.  Identification of Virus-Encoded MicroRNAs , 2004, Science.

[137]  Howard Y. Chang,et al.  Genomewide view of gene silencing by small interfering RNAs , 2003, Proceedings of the National Academy of Sciences of the United States of America.

[138]  Catherine Dargemont,et al.  Minihelix-containing RNAs Mediate Exportin-5-dependent Nuclear Export of the Double-stranded RNA-binding Protein ILF3* , 2004, Journal of Biological Chemistry.

[139]  E. Kruithof,et al.  Short-term cytotoxic effects and long-term instability of RNAi delivered using lentiviral vectors , 2004, BMC Molecular Biology.

[140]  S. Ding,et al.  Induction and Suppression of RNA Silencing by an Animal Virus , 2002, Science.

[141]  W. Dougherty,et al.  Untranslatable transcripts of the tobacco etch virus coat protein gene sequence can interfere with tobacco etch virus replication in transgenic plants and protoplasts. , 1992, Virology.

[142]  M. Matzke,et al.  Production of aberrant promoter transcripts contributes to methylation and silencing of unlinked homologous promoters in trans , 1999, The EMBO journal.

[143]  D. Weissman,et al.  Short Interfering RNA-Mediated Inhibition of Herpes Simplex Virus Type 1 Gene Expression and Function during Infection of Human Keratinocytes , 2004, Journal of Virology.

[144]  C. Kidner,et al.  Spatially restricted microRNA directs leaf polarity through ARGONAUTE1 , 2004, Nature.

[145]  W. Filipowicz,et al.  Specific interference with gene expression induced by long, double-stranded RNA in mouse embryonal teratocarcinoma cell lines , 2001, Proceedings of the National Academy of Sciences of the United States of America.

[146]  R. Kirkman,et al.  Inhibition of human immunodeficiency virus type 1 replication by siRNA targeted to the highly conserved primer binding site. , 2004, Virology.

[147]  N. Miyano-Kurosaki,et al.  Specific HIV-1 env gene silencing by small interfering RNAs in human peripheral blood mononuclear cells , 2003, Gene Therapy.

[148]  H. Giladi,et al.  Small interfering RNA inhibits hepatitis B virus replication in mice. , 2003, Molecular therapy : the journal of the American Society of Gene Therapy.

[149]  B. Cullen,et al.  Adenovirus VA1 Noncoding RNA Can Inhibit Small Interfering RNA and MicroRNA Biogenesis , 2004, Journal of Virology.

[150]  C. Rice,et al.  Clearance of replicating hepatitis C virus replicon RNAs in cell culture by small interfering RNAs , 2002, Proceedings of the National Academy of Sciences of the United States of America.

[151]  H. L. Sänger,et al.  RNA-directed de novo methylation of genomic sequences in plants , 1994, Cell.

[152]  L. Lim,et al.  An Abundant Class of Tiny RNAs with Probable Regulatory Roles in Caenorhabditis elegans , 2001, Science.

[153]  B. Heinz,et al.  Small interfering RNA molecules as potential anti-human rhinovirus agents: in vitro potency, specificity, and mechanism. , 2004, Antiviral research.

[154]  Phillip D Zamore,et al.  The RNA-Induced Silencing Complex Is a Mg2+-Dependent Endonuclease , 2004, Current Biology.

[155]  G. Dreyfuss,et al.  Numerous microRNPs in neuronal cells containing novel microRNAs. , 2003, RNA.

[156]  M. Manns,et al.  Inhibition of hepatitis B virus replication in vivo by nucleoside analogues and siRNA , 2015 .

[157]  M. Houghton,et al.  Small Interfering RNA-Mediated Inhibition of Hepatitis C Virus Replication in the Human Hepatoma Cell Line Huh-7 , 2003, Journal of Virology.

[158]  U. Kutay,et al.  Nuclear Export of MicroRNA Precursors , 2004, Science.

[159]  D. Barford,et al.  Crystal structure of a PIWI protein suggests mechanisms for siRNA recognition and slicer activity , 2004, The EMBO journal.

[160]  J. Lieberman,et al.  siRNA-directed inhibition of HIV-1 infection , 2002, Nature Medicine.

[161]  L. Vaca,et al.  Using Double-stranded RNA to Prevent in Vitro and in Vivo Viral Infections by Recombinant Baculovirus* , 2003, Journal of Biological Chemistry.

[162]  C. Hagemeier,et al.  Inhibition of human cytomegalovirus replication by small interfering RNAs. , 2004, The Journal of general virology.

[163]  P. Waterhouse,et al.  On the role of RNA silencing in the pathogenicity and evolution of viroids and viral satellites. , 2004, Proceedings of the National Academy of Sciences of the United States of America.

[164]  John Rossi,et al.  Inhibition of HIV-1 by lentiviral vector-transduced siRNAs in T lymphocytes differentiated in SCID-hu mice and CD34+ progenitor cell-derived macrophages. , 2003, Molecular therapy : the journal of the American Society of Gene Therapy.

[165]  T. Masuda,et al.  Expression of small hairpin RNA by lentivirus-based vector confers efficient and stable gene-suppression of HIV-1 on human cells including primary non-dividing cells. , 2004, Microbes and infection.

[166]  S. Barik,et al.  Inhibition of respiratory viruses by nasally administered siRNA , 2005, Nature Medicine.

[167]  M. Stevenson,et al.  Modulation of HIV-1 replication by RNA interference , 2002, Nature.

[168]  M. Wiznerowicz,et al.  Lentivirus-mediated Rna Interference of Dc-sign Expression Inhibits Human Immunodeficiency Virus Transmission from Dendritic Cells to T Cells in the Early Events of Human Immunodeficiency Virus Type 1 (hiv-1) Infection, Immature Dendritic Cells (dcs) Expressing the Dc-specific Intercellular Adhesion , 2022 .

[169]  Javier F. Palatnik,et al.  Control of leaf morphogenesis by microRNAs , 2003, Nature.

[170]  Z. Adelman,et al.  Sindbis virus‐induced silencing of dengue viruses in mosquitoes , 2001, Insect molecular biology.

[171]  Y. Stram,et al.  Inhibition of foot-and-mouth disease virus replication by small interfering RNA. , 2004, The Journal of general virology.

[172]  M. Cairns,et al.  Expressing functional siRNAs in mammalian cells using convergent transcription , 2003, BMC biotechnology.

[173]  B. Bass,et al.  A Role for the RNase III Enzyme DCR-1 in RNA Interference and Germ Line Development in Caenorhabditis elegans , 2001, Science.

[174]  Zuoshang Xu,et al.  An enhanced U6 promoter for synthesis of short hairpin RNA. , 2003, Nucleic acids research.

[175]  V. Kim,et al.  MicroRNA maturation: stepwise processing and subcellular localization , 2002, The EMBO journal.

[176]  R. Kurth,et al.  Inhibition of porcine endogenous retroviruses by RNA interference: increasing the safety of xenotransplantation. , 2004, Virology.

[177]  E. De Clercq,et al.  Selective inhibition of hepatitis B virus replication by RNA interference. , 2003, Biochemical and biophysical research communications.

[178]  B. Reinhart,et al.  The 21-nucleotide let-7 RNA regulates developmental timing in Caenorhabditis elegans , 2000, Nature.

[179]  H. Kung,et al.  Suppression of Epstein-Barr virus-encoded latent membrane protein-1 by RNA interference inhibits the metastatic potential of nasopharyngeal carcinoma cells. , 2004, Biochemical and biophysical research communications.

[180]  V. Slepushkin,et al.  Safe two‐plasmid production for the first clinical lentivirus vector that achieves >99% transduction in primary cells using a one‐step protocol , 2004, The journal of gene medicine.

[181]  Catherine H. Wu,et al.  Inhibition of HBV replication by siRNA in a stable HBV‐producing cell line , 2003, Hepatology.

[182]  K. Ikuta,et al.  RNA Interference Directed against Poly(ADP-Ribose) Polymerase 1 Efficiently Suppresses Human Immunodeficiency Virus Type 1 Replication in Human Cells , 2004, Journal of Virology.

[183]  R. Russell,et al.  bantam Encodes a Developmentally Regulated microRNA that Controls Cell Proliferation and Regulates the Proapoptotic Gene hid in Drosophila , 2003, Cell.

[184]  K. Morris,et al.  Small Interfering RNA-Induced Transcriptional Gene Silencing in Human Cells , 2004, Science.

[185]  H. Ruohola-Baker,et al.  Maelstrom, a Drosophila spindle-class gene, encodes a protein that colocalizes with Vasa and RDE1/AGO1 homolog, Aubergine, in nuage , 2003, Development.

[186]  Mark E. Davis,et al.  Lack of interferon response in animals to naked siRNAs , 2004, Nature Biotechnology.

[187]  A. Caudy,et al.  Role for a bidentate ribonuclease in the initiation step of RNA interference , 2001 .

[188]  G. Hannon,et al.  Inducible, reversible, and stable RNA interference in mammalian cells , 2004, Proceedings of the National Academy of Sciences of the United States of America.

[189]  S. Goodbourn,et al.  Interferons: cell signalling, immune modulation, antiviral response and virus countermeasures. , 2000, The Journal of general virology.

[190]  V. Kim,et al.  The nuclear RNase III Drosha initiates microRNA processing , 2003, Nature.

[191]  J. Coffin,et al.  Linkage disequilibrium test implies a large effective population number for HIV in vivo. , 1999, Proceedings of the National Academy of Sciences of the United States of America.

[192]  E. Lai.  Predicting and validating microRNA targets , 2004, Genome Biology.

[193]  Ralf Bartenschlager,et al.  Alternative Approaches for Efficient Inhibition of Hepatitis C Virus RNA Replication by Small Interfering RNAs , 2004, Journal of Virology.

[194]  G. Hannon,et al.  Crystal Structure of Argonaute and Its Implications for RISC Slicer Activity , 2004, Science.

[195]  K. Czaplinski,et al.  Exportin 5 is a RanGTP-dependent dsRNA-binding protein that mediates nuclear export of pre-miRNAs. , 2004, RNA.

[196]  S. Jayasena,et al.  Functional siRNAs and miRNAs Exhibit Strand Bias , 2003, Cell.

[197]  P. Zamore,et al.  A Protein Sensor for siRNA Asymmetry , 2004, Science.

[198]  F. Chisari,et al.  Interference of hepatitis C virus RNA replication by short interfering RNAs , 2003, Proceedings of the National Academy of Sciences of the United States of America.