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Current IssueEditorial BoardOnline First ArticlesArchive

volume 34 issue 1 (march 2013) : 62-70

RNA INTERFERENCE TECHNOLOGY: IMPLICATIONS AND APPLICATIONS-A REVIEW

S
Sanjeev Singh
I
Indrajit Ganguli
1National Bureau of Animal Genetics Resources, Karnal-132 001, India

  • Submitted|

  • First Online |

  • doi

Cite article:- Singh Sanjeev, Ganguli Indrajit (2025). RNA INTERFERENCE TECHNOLOGY: IMPLICATIONS AND APPLICATIONS-A REVIEW. Agricultural Reviews. 34(1): 62-70. doi: .

ABSTRACT

RNA interference is a phenomenon in which the introduction of double-stranded RNA (dsRNA) into a diverse range of organisms and cell types causes degradation of the complementary mRNA. ShRNA, siRNA and micro RNA are the three types of small interfering RNA involved in the gene silencing mechanism. This mechanism is triggered by double stranded RNA, which is cleaved by endoribonucleases, Drosha and Dicer to produce short RNA (si RNA) duplexes of 21-25 nt in length. The siRNA is further processed such that one of the two strands, (the passenger strand) is destroyed, while the other strand (the guide strand or antisense strand) complexes with multiple proteins to form the RNA- induced silencing complex (RISC). It is the RISC that mediates sequence specific gene silencing by recognizing the m- RNA containing sequences complimentarily with the guide strand of the siRNA. Keeping in view the importance of this highly specific DNA sequence knock down mechanism and the diverse applications of this technology, this review describes the designing of siRNAs, allele specific RNA interference, inhibition of hepatitis B Virus replication by various RNAi constructs, applications in  livestock and agricultural species, si RNA therapy, si RNA delivery systems and the challenges associated with it like immune stimulation and offtargeting effect, which should be obliterated in order to enhance the efficiency of this technology.

REFERENCES

  1. Bernstein, E; Caudy, A.A., Hammond, S.M., and Hannon, G.J. (2001). Role for a bidentate ribonuclease in the initiation step of RNA interference. Nature 409: 363-366.
  2. Carreno, V., Castillo, I., Molina, J., Porres, J. C. and Bartolome, J. (1992). Long-term follow-up of hepatitis B chronic carriers who responded to interferon therapy. J Hepatol 15: 102–106.
  3. Davuluri GR, van Tuinen A, Fraser PD, Manfredonia A, Newman R, Burgess D, Brummell DA, King SR, Palys J and Uhlig J, (2005) Fruit-specific RNAi-mediated suppression of DET1 enhances carotenoid and flavonoid content in tomatoes. Nature Biotechnology 23: 890–895.
  4. de los Santos, T., Wu, Q., de Avila Botton, S., and Grubman, M. J. (2005). Short hairpin RNA targeted to the highly conserved 2B non-structural protein coding region inhibits replication of multiple serotypes of foot-and-mouth disease virus. Virology 335, 222–231. doi:10.1016/ J.VIROL.2005.03.001
  5. Denning, C., Burl, S., Ainslie, A., Bracken, J. and Dinnyes, A.(2001). Deletion of the á(1,3)galactosyl transferase (GGTA1) gene and the prion protein (PrP) gene in sheep. Nat. Biotechnol. 19:559–562. doi:10.1038/89313.Dienstag, J. L., Cianciara, J., Karayalcin, S., Kowdley, K. V., Willems, B., Plisek, S., Woessner, M., Gardner, S. and Schiff, E. (2003). Durability of serologic response after lamivudine treatment of chronic hepatitis B. Hepatology 37: 748–755.
  6. Dykxhoorn, D. M., Schlehuber, L.D., London I.M. and Lieberman J. (2006). Determination of specific RNA interference- mediated silencing of human beta-globin alleles differing by a single nucleotide polymorphism. PNAS 103(15): 5953-5958.
  7. Elbashir, S.M., Lendeckel, W., and Tuschl, T. (2001). RNA interference mechanism is mediated by 21- and 22- nt RNAs. Genes Dev 15:188-200.
  8. Ellington, A.D. and Szostak, J.W. In vitro selection of RNA molecules that bind specific ligands. Nature 1990, 346, 818–822.
  9. Golzio, M., Mazzolini, L., Ledoux, A., Paganin, A., Izard, M., Hellaudais, L., Bieth, A., Pillaire, M.J., Cazaux, C., Hoffmann, J.S., Couderc, B. and Teissie. J. (2007). In vivo gene silencing in solid tumors by targeted electrically mediated siRNA delivery. Gene Ther. 14:752–759.
  10. Halder, J., Kamat, A.A., Jr Landen, C.N., Han, L.Y., Lutgendorf, S.K., Lin, Y.G., Merritt, W.M., Jennings, N.B., Chavez-Reyes, A., Coleman, R.L., Gershenson, D.M., Schmandt, R., Cole, S.W., Lopez-Berestein, G. and A.K. Sood, (2006). Focal adhesion kinase targeting using in vivo short interfering RNA delivery in neutral liposomes for ovarian carcinoma therapy. Clin. Cancer Res. 12:4916–4924.
  11. He, W., Liu, T., Chen, Y.X. , Cheng, D.J., Li, X.R., Xiao, Y. and Feng, Y.L. (2008) Calcium carbonate nanoparticle delivering vascular endothelial growth factor-C siRNA effectively inhibits lymphangiogenesis and growth of gastric cancer in vivo. Cancer Gene Ther. 15 193–202.
  12. Kahana, R., Kuznetzova, L., Rogel, A., Shemesh, M., Hai, D., Yadin, H., and Stram, Y. (2004). Inhibition of foot-and- mouth disease virus replication by small interfering RNA. J. Gen. Virol. 85, 3213–3217. doi:10.1099/VIR.0.80133-0
  13. Lai, C., Ching, C., Tung, A. K., Li, E., Young, J., Hill, A., Wong, B. C., Dent, J. and Wu, P. (1997). Lamivudine is effective in suppressing hepatitis B virus DNA in Chinese hepatitis B surface antigen carriers: a placebo-controlled trial. Hepatology 25: 241–244.
  14. Li, G., Jiang, P., Li,Y.,Wang, X., Huang, J., Bai, J., Cao, J.,Wu, B., Chen, N., and Zeshan, B. (2009). Inhibition of porcine reproductive and respiratory syndrome virus replication by adenovirus-mediated RNA interference both in porcine alveolar macrophages and swine. Antiviral Res. 82, 157–165. doi:10.1016/J.ANTIVIRAL.2009.02.202
  15. Li, S.D., Chen, Y.C., Hackett, M.J., and Huang, L. (2008). Tumor-targeted delivery of siRNA by self assemblednanoparticles, Mol. Ther. 16: 163–169.
  16. Magee,T. R.,Artaza, J.N., Ferrini, M. G.,Vernet,D., Zuniga, F. I., Cantini, L., Reisz-Porszasz, S., Rajfer, J., and Gonzalez- Cadavid, N. F. (2006). Myostatin short interfering hairpin RNA gene transfer increases skeletal muscle mass. J. Gene Med. 8, 1171–1181. doi:10.1002/JGM.946
  17. McPherron,A. C., Lawler,A.M., and Lee, S. J. (1997). Regulation of skeletal muscle mass in mice by a new TGF-beta superfamily member. Nature 387, 83–90. doi:10.1038/387083A0
  18. Morrissey, D.V., Blanchard, K., Shaw, L., Jensen, K., Lockridge, J.A., Dickinson, B., McSwiggen, J.A.., Vargeese, C., Bowman, K., Shaffer, C.S., Polisky, B.A. and Zinnen, S. (2005). Activity of stabilized short interfering RNA in a mouse model of hepatitis B virus replication. Hepatology 41:1349–1356.
  19. Peng, J., Zhao, Y, Mai J., Pang, W.K., Wei, X., Zhang, P. and Xu Y. (2005). Inhibition of hepatitis B virus replication by various RNAi constructs and their pharmacodynamic properties. J. General Viro. 86: 3227-3234.
  20. Pille, J.Y. Li, H. Blot, E. Bertrand, J.R. Pritchard, L.L. Opolon, P. Maksimenko, A. Lu, H.Vannier, J.P., Soria, J,. Malvy, C. and Soria, C. (2006). Intravenous delivery of anti-RhoA small interfering RNA loaded in nanoparticles of chitosan in mice: safety and efficacy in xenografted aggressive breast cancer. Hum. Gene Ther. 17: 1019–1026.
  21. Ramsoondar, J., Vaught, T., Ball, S., Mendicino, M., Monahan, J., Jobst, P., Vance, A., Duncan, J., Wells, K., and Ayares, D. (2009). Production of transgenic pigs that express porcine endogenous retrovirus small interfering RNAs. Xenotransplantation 16, 164–180. doi:10.1111/J.1399-3089.2009.00525.X
  22. Regina A, Bird A, Topping D, Bowden S, Freeman J, Barsby T, Kosar-Hashemi B, Li Z, Rahman S. and Morell M (2006). High-amylose wheat generated by RNA interference improves indices of large-bowel health in rats. Procee. National Acad. Scie. USA 103: 3546–3551.
  23. Reynolds, A., Leake, D., Boese, Q., Scaringe, S., Marshall, W.S. and Khvorova A. Rational siRNA design for RNA interference. (2004). Nature Biotechnol. 22(3):326-330.
  24. Richt, J. A., Kasinathan, P., Hamir,A. N., Castilla, J. and Sathiyaseelan, T.(2007). Production of cattle lacking prion protein. Nat. Biotechnol. 25, 132–138. doi:10.1038/NBT1271
  25. Santel, A., Aleku, M. O., Keil, J., Endruschat, V., Esche, B., Durieux, K., Löffler, M., Fechtner, T., Röhl, G., Fisch, S., Dames, W., Arnold, K., Giese, A., Klippel, J. and Kaufmann, (2006). RNA interference in the mouse vascular endothelium by systemic administration of siRNA–lipoplexes for cancer therapy. Gene Ther. 13: 1360–1370.
  26. Sledz, C.A.; Holko, M.; de Veer, M.J.; Silverman, R.H. and Williams, B.R. Activation of the interferon system by short- interfering RNAs. Nat. Cell Biol. 2003, 5, 834–839.
  27. Sunil kumar G, Campbell LM, Puckhaber L, Stipanovic RD and Rathore, K.S. (2006) Engineering cottonseed for use in human nutrition by tissue-specific reduction of toxic gossypol. Proceed. National Acad. Scie. USA 103: 18054–18059.
  28. Tang G, Galili G, Zhuang X (2007) RNAi and microRNA: breakthrough technologies for the improvement of plant nutritional value and metabolic engineering. Metabolomics 3:357–369.
  29. Tipples, G. A., Ma, M. M., Fischer, K. P., Bain, V. G., Kneteman, N. M.and Tyrrell, D. L. J. (1996). Mutation in HBV RNA-dependent DNA polymerase confers resistance to lamivudine in vivo. Hepatology 24: 714–717.
  30. Tomari Y and Zamore PD (2005) Perspective: machine for RNAi Gene Dev 19:517-529
  31. Tuerk, C. and Gold, L. Systematic evolution of ligands by exponential enrichment: RNA ligands to bacteriophage T4 DNA polymerase. Science 1990, 249, 505–510.
  32. Urban-Klein, B., Werth, S., Abuharbeid, S., Czubayko, F. and Aigner, A. (2005). RNAi-mediated gene-targeting through systemic application of polyethylenimine (PEI)-complexed siRNA in vivo. Gene Ther. 12 461–466.
  33. Van Uyen N (2006) Novel approaches in plants breeding RNAi technology. Proceedings of International Workshop on Biotechnology in Agriculture 12-16. Rodoni BC, Dale JL (1999) Harding RM Characterization and expression of the coat protein-coding region of the banana bract mosaic potyvirus, development of diagnosticassays and detection of the virus in banana plants from five countries in Southeast Asia. Archives of Virology 144:1725-1737.
  34. Wang M, Abbott D and Waterhouse P M (2000) A single copy of a virus derived transgene encoding hairpin RNA gives immunity to barley yellow dwarf virus. Molecular Plant Pathology 1: 401-410.
  35. Waterhouse PM, Graham MW and Wang MB (1998) Virus resistance and gene silencing in plants can be induced by simultaneous expression of sense and antisense RNA. Proc Natl Acad Sci USA 95: 13959–13964.
  36. Yano, J., Hirabayashi, K., Nakagawa, S., Yamaguchi, T., Nogawa, M., Kashimori, I,. Naito, H., Kitagawa, H., Ishiyama, K., Ohgi, T. and Irimura, T. (2004). Antitumor activity of small interfering RNA/cationic liposome complex in mouse models of cancer. Clin. Cancer Res. 10: 7721–7726.
  37. Zang, H., Kolb, F.A., Jaskiewicz, L., Westhof, E. and Filipowicz, W. (2004). Single processing centre models for human Dicer and bacterial RNase III. Cell 118: 57-68.
  38. Zhang Y, Engelman, J and Friedlander, R.M. (2009). Allele- specific silencing of mutant huntinton’s disease gene. J. Neurochemistry 108: 82-90.
  39. Zhang, Z., Yang, X., Zhang, Y., Zeng, B., Wang, S., Zhu, T., Roden, R.B., Chen, Y. and Yang, R. (2006). Delivery of telomerase reverse transcriptase small interfering RNA in complex with positively charged single-walled carbon nanotubes suppresses tumor growth. Clin. Cancer Res. 12: 4933–4939.
  40. Zhou, K., He, H.,Wu,Y., and Duan, M. (2008). RNA interference of avian influenza virus H5N1 by inhibiting viral mRNA with siRNA expression plasmids. J. Biotechnol. 135, 140–144. doi:10.1016/J.JBIOTEC.
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    volume 34 issue 1 (march 2013) : 62-70

    RNA INTERFERENCE TECHNOLOGY: IMPLICATIONS AND APPLICATIONS-A REVIEW

    S
    Sanjeev Singh
    I
    Indrajit Ganguli
    1National Bureau of Animal Genetics Resources, Karnal-132 001, India

    • Submitted|

    • First Online |

    • doi

    Cite article:- Singh Sanjeev, Ganguli Indrajit (2025). RNA INTERFERENCE TECHNOLOGY: IMPLICATIONS AND APPLICATIONS-A REVIEW. Agricultural Reviews. 34(1): 62-70. doi: .

    ABSTRACT

    RNA interference is a phenomenon in which the introduction of double-stranded RNA (dsRNA) into a diverse range of organisms and cell types causes degradation of the complementary mRNA. ShRNA, siRNA and micro RNA are the three types of small interfering RNA involved in the gene silencing mechanism. This mechanism is triggered by double stranded RNA, which is cleaved by endoribonucleases, Drosha and Dicer to produce short RNA (si RNA) duplexes of 21-25 nt in length. The siRNA is further processed such that one of the two strands, (the passenger strand) is destroyed, while the other strand (the guide strand or antisense strand) complexes with multiple proteins to form the RNA- induced silencing complex (RISC). It is the RISC that mediates sequence specific gene silencing by recognizing the m- RNA containing sequences complimentarily with the guide strand of the siRNA. Keeping in view the importance of this highly specific DNA sequence knock down mechanism and the diverse applications of this technology, this review describes the designing of siRNAs, allele specific RNA interference, inhibition of hepatitis B Virus replication by various RNAi constructs, applications in  livestock and agricultural species, si RNA therapy, si RNA delivery systems and the challenges associated with it like immune stimulation and offtargeting effect, which should be obliterated in order to enhance the efficiency of this technology.

    REFERENCES

    1. Bernstein, E; Caudy, A.A., Hammond, S.M., and Hannon, G.J. (2001). Role for a bidentate ribonuclease in the initiation step of RNA interference. Nature 409: 363-366.
    2. Carreno, V., Castillo, I., Molina, J., Porres, J. C. and Bartolome, J. (1992). Long-term follow-up of hepatitis B chronic carriers who responded to interferon therapy. J Hepatol 15: 102–106.
    3. Davuluri GR, van Tuinen A, Fraser PD, Manfredonia A, Newman R, Burgess D, Brummell DA, King SR, Palys J and Uhlig J, (2005) Fruit-specific RNAi-mediated suppression of DET1 enhances carotenoid and flavonoid content in tomatoes. Nature Biotechnology 23: 890–895.
    4. de los Santos, T., Wu, Q., de Avila Botton, S., and Grubman, M. J. (2005). Short hairpin RNA targeted to the highly conserved 2B non-structural protein coding region inhibits replication of multiple serotypes of foot-and-mouth disease virus. Virology 335, 222–231. doi:10.1016/ J.VIROL.2005.03.001
    5. Denning, C., Burl, S., Ainslie, A., Bracken, J. and Dinnyes, A.(2001). Deletion of the á(1,3)galactosyl transferase (GGTA1) gene and the prion protein (PrP) gene in sheep. Nat. Biotechnol. 19:559–562. doi:10.1038/89313.Dienstag, J. L., Cianciara, J., Karayalcin, S., Kowdley, K. V., Willems, B., Plisek, S., Woessner, M., Gardner, S. and Schiff, E. (2003). Durability of serologic response after lamivudine treatment of chronic hepatitis B. Hepatology 37: 748–755.
    6. Dykxhoorn, D. M., Schlehuber, L.D., London I.M. and Lieberman J. (2006). Determination of specific RNA interference- mediated silencing of human beta-globin alleles differing by a single nucleotide polymorphism. PNAS 103(15): 5953-5958.
    7. Elbashir, S.M., Lendeckel, W., and Tuschl, T. (2001). RNA interference mechanism is mediated by 21- and 22- nt RNAs. Genes Dev 15:188-200.
    8. Ellington, A.D. and Szostak, J.W. In vitro selection of RNA molecules that bind specific ligands. Nature 1990, 346, 818–822.
    9. Golzio, M., Mazzolini, L., Ledoux, A., Paganin, A., Izard, M., Hellaudais, L., Bieth, A., Pillaire, M.J., Cazaux, C., Hoffmann, J.S., Couderc, B. and Teissie. J. (2007). In vivo gene silencing in solid tumors by targeted electrically mediated siRNA delivery. Gene Ther. 14:752–759.
    10. Halder, J., Kamat, A.A., Jr Landen, C.N., Han, L.Y., Lutgendorf, S.K., Lin, Y.G., Merritt, W.M., Jennings, N.B., Chavez-Reyes, A., Coleman, R.L., Gershenson, D.M., Schmandt, R., Cole, S.W., Lopez-Berestein, G. and A.K. Sood, (2006). Focal adhesion kinase targeting using in vivo short interfering RNA delivery in neutral liposomes for ovarian carcinoma therapy. Clin. Cancer Res. 12:4916–4924.
    11. He, W., Liu, T., Chen, Y.X. , Cheng, D.J., Li, X.R., Xiao, Y. and Feng, Y.L. (2008) Calcium carbonate nanoparticle delivering vascular endothelial growth factor-C siRNA effectively inhibits lymphangiogenesis and growth of gastric cancer in vivo. Cancer Gene Ther. 15 193–202.
    12. Kahana, R., Kuznetzova, L., Rogel, A., Shemesh, M., Hai, D., Yadin, H., and Stram, Y. (2004). Inhibition of foot-and- mouth disease virus replication by small interfering RNA. J. Gen. Virol. 85, 3213–3217. doi:10.1099/VIR.0.80133-0
    13. Lai, C., Ching, C., Tung, A. K., Li, E., Young, J., Hill, A., Wong, B. C., Dent, J. and Wu, P. (1997). Lamivudine is effective in suppressing hepatitis B virus DNA in Chinese hepatitis B surface antigen carriers: a placebo-controlled trial. Hepatology 25: 241–244.
    14. Li, G., Jiang, P., Li,Y.,Wang, X., Huang, J., Bai, J., Cao, J.,Wu, B., Chen, N., and Zeshan, B. (2009). Inhibition of porcine reproductive and respiratory syndrome virus replication by adenovirus-mediated RNA interference both in porcine alveolar macrophages and swine. Antiviral Res. 82, 157–165. doi:10.1016/J.ANTIVIRAL.2009.02.202
    15. Li, S.D., Chen, Y.C., Hackett, M.J., and Huang, L. (2008). Tumor-targeted delivery of siRNA by self assemblednanoparticles, Mol. Ther. 16: 163–169.
    16. Magee,T. R.,Artaza, J.N., Ferrini, M. G.,Vernet,D., Zuniga, F. I., Cantini, L., Reisz-Porszasz, S., Rajfer, J., and Gonzalez- Cadavid, N. F. (2006). Myostatin short interfering hairpin RNA gene transfer increases skeletal muscle mass. J. Gene Med. 8, 1171–1181. doi:10.1002/JGM.946
    17. McPherron,A. C., Lawler,A.M., and Lee, S. J. (1997). Regulation of skeletal muscle mass in mice by a new TGF-beta superfamily member. Nature 387, 83–90. doi:10.1038/387083A0
    18. Morrissey, D.V., Blanchard, K., Shaw, L., Jensen, K., Lockridge, J.A., Dickinson, B., McSwiggen, J.A.., Vargeese, C., Bowman, K., Shaffer, C.S., Polisky, B.A. and Zinnen, S. (2005). Activity of stabilized short interfering RNA in a mouse model of hepatitis B virus replication. Hepatology 41:1349–1356.
    19. Peng, J., Zhao, Y, Mai J., Pang, W.K., Wei, X., Zhang, P. and Xu Y. (2005). Inhibition of hepatitis B virus replication by various RNAi constructs and their pharmacodynamic properties. J. General Viro. 86: 3227-3234.
    20. Pille, J.Y. Li, H. Blot, E. Bertrand, J.R. Pritchard, L.L. Opolon, P. Maksimenko, A. Lu, H.Vannier, J.P., Soria, J,. Malvy, C. and Soria, C. (2006). Intravenous delivery of anti-RhoA small interfering RNA loaded in nanoparticles of chitosan in mice: safety and efficacy in xenografted aggressive breast cancer. Hum. Gene Ther. 17: 1019–1026.
    21. Ramsoondar, J., Vaught, T., Ball, S., Mendicino, M., Monahan, J., Jobst, P., Vance, A., Duncan, J., Wells, K., and Ayares, D. (2009). Production of transgenic pigs that express porcine endogenous retrovirus small interfering RNAs. Xenotransplantation 16, 164–180. doi:10.1111/J.1399-3089.2009.00525.X
    22. Regina A, Bird A, Topping D, Bowden S, Freeman J, Barsby T, Kosar-Hashemi B, Li Z, Rahman S. and Morell M (2006). High-amylose wheat generated by RNA interference improves indices of large-bowel health in rats. Procee. National Acad. Scie. USA 103: 3546–3551.
    23. Reynolds, A., Leake, D., Boese, Q., Scaringe, S., Marshall, W.S. and Khvorova A. Rational siRNA design for RNA interference. (2004). Nature Biotechnol. 22(3):326-330.
    24. Richt, J. A., Kasinathan, P., Hamir,A. N., Castilla, J. and Sathiyaseelan, T.(2007). Production of cattle lacking prion protein. Nat. Biotechnol. 25, 132–138. doi:10.1038/NBT1271
    25. Santel, A., Aleku, M. O., Keil, J., Endruschat, V., Esche, B., Durieux, K., Löffler, M., Fechtner, T., Röhl, G., Fisch, S., Dames, W., Arnold, K., Giese, A., Klippel, J. and Kaufmann, (2006). RNA interference in the mouse vascular endothelium by systemic administration of siRNA–lipoplexes for cancer therapy. Gene Ther. 13: 1360–1370.
    26. Sledz, C.A.; Holko, M.; de Veer, M.J.; Silverman, R.H. and Williams, B.R. Activation of the interferon system by short- interfering RNAs. Nat. Cell Biol. 2003, 5, 834–839.
    27. Sunil kumar G, Campbell LM, Puckhaber L, Stipanovic RD and Rathore, K.S. (2006) Engineering cottonseed for use in human nutrition by tissue-specific reduction of toxic gossypol. Proceed. National Acad. Scie. USA 103: 18054–18059.
    28. Tang G, Galili G, Zhuang X (2007) RNAi and microRNA: breakthrough technologies for the improvement of plant nutritional value and metabolic engineering. Metabolomics 3:357–369.
    29. Tipples, G. A., Ma, M. M., Fischer, K. P., Bain, V. G., Kneteman, N. M.and Tyrrell, D. L. J. (1996). Mutation in HBV RNA-dependent DNA polymerase confers resistance to lamivudine in vivo. Hepatology 24: 714–717.
    30. Tomari Y and Zamore PD (2005) Perspective: machine for RNAi Gene Dev 19:517-529
    31. Tuerk, C. and Gold, L. Systematic evolution of ligands by exponential enrichment: RNA ligands to bacteriophage T4 DNA polymerase. Science 1990, 249, 505–510.
    32. Urban-Klein, B., Werth, S., Abuharbeid, S., Czubayko, F. and Aigner, A. (2005). RNAi-mediated gene-targeting through systemic application of polyethylenimine (PEI)-complexed siRNA in vivo. Gene Ther. 12 461–466.
    33. Van Uyen N (2006) Novel approaches in plants breeding RNAi technology. Proceedings of International Workshop on Biotechnology in Agriculture 12-16. Rodoni BC, Dale JL (1999) Harding RM Characterization and expression of the coat protein-coding region of the banana bract mosaic potyvirus, development of diagnosticassays and detection of the virus in banana plants from five countries in Southeast Asia. Archives of Virology 144:1725-1737.
    34. Wang M, Abbott D and Waterhouse P M (2000) A single copy of a virus derived transgene encoding hairpin RNA gives immunity to barley yellow dwarf virus. Molecular Plant Pathology 1: 401-410.
    35. Waterhouse PM, Graham MW and Wang MB (1998) Virus resistance and gene silencing in plants can be induced by simultaneous expression of sense and antisense RNA. Proc Natl Acad Sci USA 95: 13959–13964.
    36. Yano, J., Hirabayashi, K., Nakagawa, S., Yamaguchi, T., Nogawa, M., Kashimori, I,. Naito, H., Kitagawa, H., Ishiyama, K., Ohgi, T. and Irimura, T. (2004). Antitumor activity of small interfering RNA/cationic liposome complex in mouse models of cancer. Clin. Cancer Res. 10: 7721–7726.
    37. Zang, H., Kolb, F.A., Jaskiewicz, L., Westhof, E. and Filipowicz, W. (2004). Single processing centre models for human Dicer and bacterial RNase III. Cell 118: 57-68.
    38. Zhang Y, Engelman, J and Friedlander, R.M. (2009). Allele- specific silencing of mutant huntinton’s disease gene. J. Neurochemistry 108: 82-90.
    39. Zhang, Z., Yang, X., Zhang, Y., Zeng, B., Wang, S., Zhu, T., Roden, R.B., Chen, Y. and Yang, R. (2006). Delivery of telomerase reverse transcriptase small interfering RNA in complex with positively charged single-walled carbon nanotubes suppresses tumor growth. Clin. Cancer Res. 12: 4933–4939.
    40. Zhou, K., He, H.,Wu,Y., and Duan, M. (2008). RNA interference of avian influenza virus H5N1 by inhibiting viral mRNA with siRNA expression plasmids. J. Biotechnol. 135, 140–144. doi:10.1016/J.JBIOTEC.
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