Skip to main content
Top
Published in: Virology Journal 1/2011

Open Access 01-12-2011 | Research

Inhibition of full length Hepatitis C Virus particles of 1a genotype through small interference RNA

Authors: Muhammad Ansar, Usman Ali Ashfaq, Imran shahid, Muhammad Tahir Sarwar, Tariq Javed, Sidra Rehman, Sajida Hassan, Sheikh Riazuddin

Published in: Virology Journal | Issue 1/2011

Login to get access

Abstract

Background

Hepatitis C virus (HCV), a member of the Flaviviridae family of viruses, is a major cause of chronic hepatitis, liver cirrhosis and hepatocellular carcinoma. Currently, the only treatment available consists of a combination of Pegylated interferon alpha (INF-α) and ribavirin, but only half of the patients treated show a sufficient antiviral response. Thus there is a great need for the development of new treatments for HCV infections. RNA interference (RNAi) represents a new promising approach to develop effective antiviral drugs and has been extremely effective against HCV infection.

Results

This study was design to assess or explore the silencing effect of small interference RNAs (siRNAs) against full length HCV particles of genotype 1a. In the present study six 21-bp siRNAs were designed against different regions of HCV structural genes (Core, E1 and E2). Selected siRNAs were labeled as Csi 301, Csi 29, E1si 52, E1si 192, E2si 86 and E2si 493. Our results demonstrated that siRNAs directed against HCV core gene showed 70% reduction in viral titer in HCV infected liver cells. Moreover, siRNAs against E1 and E2 envelop genes showed a dramatic reduction in HCV viral RNA, E2si 86 exhibited 93% inhibition, while E1si 192, E2si 493 and E1si 52 showed 87%, 80%, and 66% inhibition respectively. No significant inhibition was detected in cells transfected with the negative control siRNA.

Conclusion

Our results suggested that siRNAs targeted against HCV structural genes efficiently silence full length HCV particles and provide an effective therapeutic option against HCV infection.
Appendix
Available only for authorised users
Literature
1.
go back to reference Choo QL, Kuo G, Weiner AJ, Overby LR, Bradley DW, Houghton M: Isolation of a cDNA clone derived from a blood-borne non-A, non-B viral hepatitis genome. Science 1989, 244: 359-362. 10.1126/science.2523562CrossRefPubMed Choo QL, Kuo G, Weiner AJ, Overby LR, Bradley DW, Houghton M: Isolation of a cDNA clone derived from a blood-borne non-A, non-B viral hepatitis genome. Science 1989, 244: 359-362. 10.1126/science.2523562CrossRefPubMed
2.
go back to reference Lin C, Lindenbach BD, Pragai BM, McCourt DW, Rice CM: Processing in the hepatitis C virus E2-NS2 region: identification of p7 and two distinct E2-specific products with different C termini. J Virol 1994, 68: 5063-5073.PubMedCentralPubMed Lin C, Lindenbach BD, Pragai BM, McCourt DW, Rice CM: Processing in the hepatitis C virus E2-NS2 region: identification of p7 and two distinct E2-specific products with different C termini. J Virol 1994, 68: 5063-5073.PubMedCentralPubMed
3.
go back to reference Reed KE, Rice CM: Overview of hepatitis C virus genome structure, polyprotein processing, and protein properties. Curr Top Microbiol Immunol 2000, 242: 55-84.PubMed Reed KE, Rice CM: Overview of hepatitis C virus genome structure, polyprotein processing, and protein properties. Curr Top Microbiol Immunol 2000, 242: 55-84.PubMed
4.
go back to reference Moriya K, Nakagawa K, Santa T, Shintani Y, Fujie H, Miyoshi H, Tsutsumi T, Miyazawa T, Ishibashi K, Horie T, et al.: Oxidative stress in the absence of inflammation in a mouse model for hepatitis C virus-associated hepatocarcinogenesis. Cancer Res 2001, 61: 4365-4370.PubMed Moriya K, Nakagawa K, Santa T, Shintani Y, Fujie H, Miyoshi H, Tsutsumi T, Miyazawa T, Ishibashi K, Horie T, et al.: Oxidative stress in the absence of inflammation in a mouse model for hepatitis C virus-associated hepatocarcinogenesis. Cancer Res 2001, 61: 4365-4370.PubMed
5.
go back to reference Liu Q, Tackney C, Bhat RA, Prince AM, Zhang P: Regulated processing of hepatitis C virus core protein is linked to subcellular localization. J Virol 1997, 71: 657-662.PubMedCentralPubMed Liu Q, Tackney C, Bhat RA, Prince AM, Zhang P: Regulated processing of hepatitis C virus core protein is linked to subcellular localization. J Virol 1997, 71: 657-662.PubMedCentralPubMed
7.
go back to reference Memon MI, Memon MA: Hepatitis C: an epidemiological review. J Viral Hepat 2002, 9: 84-100. 10.1046/j.1365-2893.2002.00329.xCrossRefPubMed Memon MI, Memon MA: Hepatitis C: an epidemiological review. J Viral Hepat 2002, 9: 84-100. 10.1046/j.1365-2893.2002.00329.xCrossRefPubMed
8.
go back to reference Manns MP, McHutchison JG, Gordon SC, Rustgi VK, Shiffman M, Reindollar R, Goodman ZD, Koury K, Ling M, Albrecht JK: Peginterferon alfa-2b plus ribavirin compared with interferon alfa-2b plus ribavirin for initial treatment of chronic hepatitis C: a randomised trial. Lancet 2001, 358: 958-965. 10.1016/S0140-6736(01)06102-5CrossRefPubMed Manns MP, McHutchison JG, Gordon SC, Rustgi VK, Shiffman M, Reindollar R, Goodman ZD, Koury K, Ling M, Albrecht JK: Peginterferon alfa-2b plus ribavirin compared with interferon alfa-2b plus ribavirin for initial treatment of chronic hepatitis C: a randomised trial. Lancet 2001, 358: 958-965. 10.1016/S0140-6736(01)06102-5CrossRefPubMed
9.
10.
go back to reference Fried MW, Shiffman ML, Reddy KR, Smith C, Marinos G, Goncales FL Jr, Haussinger D, Diago M, Carosi G, Dhumeaux D, et al.: Peginterferon alfa-2a plus ribavirin for chronic hepatitis C virus infection. N Engl J Med 2002, 347: 975-982. 10.1056/NEJMoa020047CrossRefPubMed Fried MW, Shiffman ML, Reddy KR, Smith C, Marinos G, Goncales FL Jr, Haussinger D, Diago M, Carosi G, Dhumeaux D, et al.: Peginterferon alfa-2a plus ribavirin for chronic hepatitis C virus infection. N Engl J Med 2002, 347: 975-982. 10.1056/NEJMoa020047CrossRefPubMed
11.
go back to reference Fire A, Xu S, Montgomery MK, Kostas SA, Driver SE, Mello CC: Potent and specific genetic interference by double-stranded RNA in Caenorhabditis elegans. Nature 1998, 391: 806-811. 10.1038/35888CrossRefPubMed Fire A, Xu S, Montgomery MK, Kostas SA, Driver SE, Mello CC: Potent and specific genetic interference by double-stranded RNA in Caenorhabditis elegans. Nature 1998, 391: 806-811. 10.1038/35888CrossRefPubMed
12.
go back to reference Elbashir SM, Harborth J, Lendeckel W, Yalcin A, Weber K, Tuschl T: Duplexes of 21-nucleotide RNAs mediate RNA interference in cultured mammalian cells. Nature 2001, 411: 494-498. 10.1038/35078107CrossRefPubMed Elbashir SM, Harborth J, Lendeckel W, Yalcin A, Weber K, Tuschl T: Duplexes of 21-nucleotide RNAs mediate RNA interference in cultured mammalian cells. Nature 2001, 411: 494-498. 10.1038/35078107CrossRefPubMed
13.
14.
go back to reference Elbashir SM, Martinez J, Patkaniowska A, Lendeckel W, Tuschl T: Functional anatomy of siRNAs for mediating efficient RNAi in Drosophila melanogaster embryo lysate. Embo J 2001, 20: 6877-6888. 10.1093/emboj/20.23.6877PubMedCentralCrossRefPubMed Elbashir SM, Martinez J, Patkaniowska A, Lendeckel W, Tuschl T: Functional anatomy of siRNAs for mediating efficient RNAi in Drosophila melanogaster embryo lysate. Embo J 2001, 20: 6877-6888. 10.1093/emboj/20.23.6877PubMedCentralCrossRefPubMed
16.
go back to reference Cullen BR: RNA interference: antiviral defense and genetic tool. Nat Immunol 2002, 3: 597-599. 10.1038/ni0702-597CrossRefPubMed Cullen BR: RNA interference: antiviral defense and genetic tool. Nat Immunol 2002, 3: 597-599. 10.1038/ni0702-597CrossRefPubMed
18.
go back to reference Tuschl T: RNA interference and small interfering RNAs. Chembiochem 2001, 2: 239-245. 10.1002/1439-7633(20010401)2:4<239::AID-CBIC239>3.0.CO;2-RCrossRefPubMed Tuschl T: RNA interference and small interfering RNAs. Chembiochem 2001, 2: 239-245. 10.1002/1439-7633(20010401)2:4<239::AID-CBIC239>3.0.CO;2-RCrossRefPubMed
19.
go back to reference Scherer LJ, Rossi JJ: Approaches for the sequence-specific knockdown of mRNA. Nat Biotechnol 2003, 21: 1457-1465. 10.1038/nbt915CrossRefPubMed Scherer LJ, Rossi JJ: Approaches for the sequence-specific knockdown of mRNA. Nat Biotechnol 2003, 21: 1457-1465. 10.1038/nbt915CrossRefPubMed
20.
go back to reference Sahu NK, Shilakari G, Nayak A, Kohli DV: Antisense technology: a selective tool for gene expression regulation and gene targeting. Curr Pharm Biotechnol 2007, 8: 291-304. 10.2174/138920107782109985CrossRefPubMed Sahu NK, Shilakari G, Nayak A, Kohli DV: Antisense technology: a selective tool for gene expression regulation and gene targeting. Curr Pharm Biotechnol 2007, 8: 291-304. 10.2174/138920107782109985CrossRefPubMed
21.
go back to reference Randall G, Panis M, Cooper JD, Tellinghuisen TL, Sukhodolets KE, Pfeffer S, Landthaler M, Landgraf P, Kan S, Lindenbach BD, et al.: Cellular cofactors affecting hepatitis C virus infection and replication. Proc Natl Acad Sci USA 2007, 104: 12884-12889. 10.1073/pnas.0704894104PubMedCentralCrossRefPubMed Randall G, Panis M, Cooper JD, Tellinghuisen TL, Sukhodolets KE, Pfeffer S, Landthaler M, Landgraf P, Kan S, Lindenbach BD, et al.: Cellular cofactors affecting hepatitis C virus infection and replication. Proc Natl Acad Sci USA 2007, 104: 12884-12889. 10.1073/pnas.0704894104PubMedCentralCrossRefPubMed
22.
go back to reference McCaffrey AP, Meuse L, Pham TT, Conklin DS, Hannon GJ, Kay MA: RNA interference in adult mice. Nature 2002, 418: 38-39. 10.1038/418038aCrossRefPubMed McCaffrey AP, Meuse L, Pham TT, Conklin DS, Hannon GJ, Kay MA: RNA interference in adult mice. Nature 2002, 418: 38-39. 10.1038/418038aCrossRefPubMed
23.
go back to reference Kapadia SB, Brideau-Andersen A, Chisari FV: Interference of hepatitis C virus RNA replication by short interfering RNAs. Proc Natl Acad Sci USA 2003, 100: 2014-2018. 10.1073/pnas.252783999PubMedCentralCrossRefPubMed Kapadia SB, Brideau-Andersen A, Chisari FV: Interference of hepatitis C virus RNA replication by short interfering RNAs. Proc Natl Acad Sci USA 2003, 100: 2014-2018. 10.1073/pnas.252783999PubMedCentralCrossRefPubMed
24.
go back to reference Randall G, Chen L, Panis M, Fischer AK, Lindenbach BD, Sun J, Heathcote J, Rice CM, Edwards AM, McGilvray ID: Silencing of USP18 potentiates the antiviral activity of interferon against hepatitis C virus infection. Gastroenterology 2006, 131: 1584-1591. 10.1053/j.gastro.2006.08.043CrossRefPubMed Randall G, Chen L, Panis M, Fischer AK, Lindenbach BD, Sun J, Heathcote J, Rice CM, Edwards AM, McGilvray ID: Silencing of USP18 potentiates the antiviral activity of interferon against hepatitis C virus infection. Gastroenterology 2006, 131: 1584-1591. 10.1053/j.gastro.2006.08.043CrossRefPubMed
25.
go back to reference Wilson JA, Jayasena S, Khvorova A, Sabatinos S, Rodrigue-Gervais IG, Arya S, Sarangi F, Harris-Brandts M, Beaulieu S, Richardson CD: RNA interference blocks gene expression and RNA synthesis from hepatitis C replicons propagated in human liver cells. Proc Natl Acad Sci USA 2003, 100: 2783-2788. 10.1073/pnas.252758799PubMedCentralCrossRefPubMed Wilson JA, Jayasena S, Khvorova A, Sabatinos S, Rodrigue-Gervais IG, Arya S, Sarangi F, Harris-Brandts M, Beaulieu S, Richardson CD: RNA interference blocks gene expression and RNA synthesis from hepatitis C replicons propagated in human liver cells. Proc Natl Acad Sci USA 2003, 100: 2783-2788. 10.1073/pnas.252758799PubMedCentralCrossRefPubMed
26.
go back to reference Takigawa Y, Nagano-Fujii M, Deng L, Hidajat R, Tanaka M, Mizuta H, Hotta H: Suppression of hepatitis C virus replicon by RNA interference directed against the NS3 and NS5B regions of the viral genome. Microbiol Immunol 2004, 48: 591-598.CrossRefPubMed Takigawa Y, Nagano-Fujii M, Deng L, Hidajat R, Tanaka M, Mizuta H, Hotta H: Suppression of hepatitis C virus replicon by RNA interference directed against the NS3 and NS5B regions of the viral genome. Microbiol Immunol 2004, 48: 591-598.CrossRefPubMed
27.
28.
go back to reference el-Awady MK, Tabll AA, el-Abd YS, Bahgat MM, Shoeb HA, Youssef SS, Bader el-Din NG, Redwan el RM, el-Demellawy M, Omran MH, et al.: HepG2 cells support viral replication and gene expression of hepatitis C virus genotype 4 in vitro. World J Gastroenterol 2006, 12: 4836-4842.PubMedCentralPubMed el-Awady MK, Tabll AA, el-Abd YS, Bahgat MM, Shoeb HA, Youssef SS, Bader el-Din NG, Redwan el RM, el-Demellawy M, Omran MH, et al.: HepG2 cells support viral replication and gene expression of hepatitis C virus genotype 4 in vitro. World J Gastroenterol 2006, 12: 4836-4842.PubMedCentralPubMed
29.
go back to reference Mosmann T: Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays. J Immunol Methods 1983, 65: 55-63. 10.1016/0022-1759(83)90303-4CrossRefPubMed Mosmann T: Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays. J Immunol Methods 1983, 65: 55-63. 10.1016/0022-1759(83)90303-4CrossRefPubMed
30.
go back to reference Khaliq S, Khaliq SA, Zahur M, Ijaz B, Jahan S, Ansar M, Riazuddin S, Hassan S: RNAi as a new therapeutic strategy against HCV. Biotechnol Adv 28: 27-34. Khaliq S, Khaliq SA, Zahur M, Ijaz B, Jahan S, Ansar M, Riazuddin S, Hassan S: RNAi as a new therapeutic strategy against HCV. Biotechnol Adv 28: 27-34.
31.
go back to reference Caplen NJ, Parrish S, Imani F, Fire A, Morgan RA: Specific inhibition of gene expression by small double-stranded RNAs in invertebrate and vertebrate systems. Proc Natl Acad Sci USA 2001, 98: 9742-9747. 10.1073/pnas.171251798PubMedCentralCrossRefPubMed Caplen NJ, Parrish S, Imani F, Fire A, Morgan RA: Specific inhibition of gene expression by small double-stranded RNAs in invertebrate and vertebrate systems. Proc Natl Acad Sci USA 2001, 98: 9742-9747. 10.1073/pnas.171251798PubMedCentralCrossRefPubMed
32.
go back to reference Shin D, Lee H, Kim SI, Yoon Y, Kim M: Optimization of linear double-stranded RNA for the production of multiple siRNAs targeting hepatitis C virus. Rna 2009, 15: 898-910. 10.1261/rna.1268209PubMedCentralCrossRefPubMed Shin D, Lee H, Kim SI, Yoon Y, Kim M: Optimization of linear double-stranded RNA for the production of multiple siRNAs targeting hepatitis C virus. Rna 2009, 15: 898-910. 10.1261/rna.1268209PubMedCentralCrossRefPubMed
33.
go back to reference Bitko V, Musiyenko A, Shulyayeva O, Barik S: Inhibition of respiratory viruses by nasally administered siRNA. Nat Med 2005, 11: 50-55. 10.1038/nm1164CrossRefPubMed Bitko V, Musiyenko A, Shulyayeva O, Barik S: Inhibition of respiratory viruses by nasally administered siRNA. Nat Med 2005, 11: 50-55. 10.1038/nm1164CrossRefPubMed
34.
go back to reference Brummelkamp TR, Bernards R, Agami R: A system for stable expression of short interfering RNAs in mammalian cells. Science 2002, 296: 550-553. 10.1126/science.1068999CrossRefPubMed Brummelkamp TR, Bernards R, Agami R: A system for stable expression of short interfering RNAs in mammalian cells. Science 2002, 296: 550-553. 10.1126/science.1068999CrossRefPubMed
35.
go back to reference Prabhu R, Vittal P, Yin Q, Flemington E, Garry R, Robichaux WH, Dash S: Small interfering RNA effectively inhibits protein expression and negative strand RNA synthesis from a full-length hepatitis C virus clone. J Med Virol 2005, 76: 511-519. 10.1002/jmv.20391CrossRefPubMed Prabhu R, Vittal P, Yin Q, Flemington E, Garry R, Robichaux WH, Dash S: Small interfering RNA effectively inhibits protein expression and negative strand RNA synthesis from a full-length hepatitis C virus clone. J Med Virol 2005, 76: 511-519. 10.1002/jmv.20391CrossRefPubMed
36.
go back to reference Wilson JA, Richardson CD: Hepatitis C virus replicons escape RNA interference induced by a short interfering RNA directed against the NS5b coding region. J Virol 2005, 79: 7050-7058. 10.1128/JVI.79.11.7050-7058.2005PubMedCentralCrossRefPubMed Wilson JA, Richardson CD: Hepatitis C virus replicons escape RNA interference induced by a short interfering RNA directed against the NS5b coding region. J Virol 2005, 79: 7050-7058. 10.1128/JVI.79.11.7050-7058.2005PubMedCentralCrossRefPubMed
37.
go back to reference Watanabe T, Umehara T, Yasui F, Nakagawa S, Yano J, Ohgi T, Sonoke S, Satoh K, Inoue K, Yoshiba M, Kohara M: Liver target delivery of small interfering RNA to the HCV gene by lactosylated cationic liposome. J Hepatol 2007, 47: 744-750. 10.1016/j.jhep.2007.06.015CrossRefPubMed Watanabe T, Umehara T, Yasui F, Nakagawa S, Yano J, Ohgi T, Sonoke S, Satoh K, Inoue K, Yoshiba M, Kohara M: Liver target delivery of small interfering RNA to the HCV gene by lactosylated cationic liposome. J Hepatol 2007, 47: 744-750. 10.1016/j.jhep.2007.06.015CrossRefPubMed
38.
go back to reference Seo MY, Abrignani S, Houghton M, Han JH: Small interfering RNA-mediated inhibition of hepatitis C virus replication in the human hepatoma cell line Huh-7. J Virol 2003, 77: 810-812. 10.1128/JVI.77.1.810-812.2003PubMedCentralCrossRefPubMed Seo MY, Abrignani S, Houghton M, Han JH: Small interfering RNA-mediated inhibition of hepatitis C virus replication in the human hepatoma cell line Huh-7. J Virol 2003, 77: 810-812. 10.1128/JVI.77.1.810-812.2003PubMedCentralCrossRefPubMed
39.
go back to reference Yokota T, Sakamoto N, Enomoto N, Tanabe Y, Miyagishi M, Maekawa S, Yi L, Kurosaki M, Taira K, Watanabe M, Mizusawa H: Inhibition of intracellular hepatitis C virus replication by synthetic and vector-derived small interfering RNAs. EMBO Rep 2003, 4: 602-608. 10.1038/sj.embor.embor840PubMedCentralCrossRefPubMed Yokota T, Sakamoto N, Enomoto N, Tanabe Y, Miyagishi M, Maekawa S, Yi L, Kurosaki M, Taira K, Watanabe M, Mizusawa H: Inhibition of intracellular hepatitis C virus replication by synthetic and vector-derived small interfering RNAs. EMBO Rep 2003, 4: 602-608. 10.1038/sj.embor.embor840PubMedCentralCrossRefPubMed
40.
go back to reference Kim M, Shin D, Kim SI, Park M: Inhibition of hepatitis C virus gene expression by small interfering RNAs using a tri-cistronic full-length viral replicon and a transient mouse model. Virus Res 2006, 122: 1-10. 10.1016/j.virusres.2006.05.003CrossRefPubMed Kim M, Shin D, Kim SI, Park M: Inhibition of hepatitis C virus gene expression by small interfering RNAs using a tri-cistronic full-length viral replicon and a transient mouse model. Virus Res 2006, 122: 1-10. 10.1016/j.virusres.2006.05.003CrossRefPubMed
41.
go back to reference Liu M, Ding H, Zhao P, Qin ZL, Gao J, Cao MM, Luan J, Wu WB, Qi ZT: RNA interference effectively inhibits mRNA accumulation and protein expression of hepatitis C virus core and E2 genes in human cells. Biosci Biotechnol Biochem 2006, 70: 2049-2055. 10.1271/bbb.60001CrossRefPubMed Liu M, Ding H, Zhao P, Qin ZL, Gao J, Cao MM, Luan J, Wu WB, Qi ZT: RNA interference effectively inhibits mRNA accumulation and protein expression of hepatitis C virus core and E2 genes in human cells. Biosci Biotechnol Biochem 2006, 70: 2049-2055. 10.1271/bbb.60001CrossRefPubMed
42.
go back to reference Zhong J, Gastaminza P, Cheng G, Kapadia S, Kato T, Burton DR, Wieland SF, Uprichard SL, Wakita T, Chisari FV: Robust hepatitis C virus infection in vitro. Proc Natl Acad Sci USA 2005, 102: 9294-9299. 10.1073/pnas.0503596102PubMedCentralCrossRefPubMed Zhong J, Gastaminza P, Cheng G, Kapadia S, Kato T, Burton DR, Wieland SF, Uprichard SL, Wakita T, Chisari FV: Robust hepatitis C virus infection in vitro. Proc Natl Acad Sci USA 2005, 102: 9294-9299. 10.1073/pnas.0503596102PubMedCentralCrossRefPubMed
43.
go back to reference Lazaro CA, Chang M, Tang W, Campbell J, Sullivan DG, Gretch DR, Corey L, Coombs RW, Fausto N: Hepatitis C virus replication in transfected and serum-infected cultured human fetal hepatocytes. Am J Pathol 2007, 170: 478-489. 10.2353/ajpath.2007.060789PubMedCentralCrossRefPubMed Lazaro CA, Chang M, Tang W, Campbell J, Sullivan DG, Gretch DR, Corey L, Coombs RW, Fausto N: Hepatitis C virus replication in transfected and serum-infected cultured human fetal hepatocytes. Am J Pathol 2007, 170: 478-489. 10.2353/ajpath.2007.060789PubMedCentralCrossRefPubMed
44.
go back to reference Buck M: Direct infection and replication of naturally occurring hepatitis C virus genotypes 1, 2, 3 and 4 in normal human hepatocyte cultures. PLoS One 2008, 3: e2660. 10.1371/journal.pone.0002660PubMedCentralCrossRefPubMed Buck M: Direct infection and replication of naturally occurring hepatitis C virus genotypes 1, 2, 3 and 4 in normal human hepatocyte cultures. PLoS One 2008, 3: e2660. 10.1371/journal.pone.0002660PubMedCentralCrossRefPubMed
Metadata
Title
Inhibition of full length Hepatitis C Virus particles of 1a genotype through small interference RNA
Authors
Muhammad Ansar
Usman Ali Ashfaq
Imran shahid
Muhammad Tahir Sarwar
Tariq Javed
Sidra Rehman
Sajida Hassan
Sheikh Riazuddin
Publication date
01-12-2011
Publisher
BioMed Central
Published in
Virology Journal / Issue 1/2011
Electronic ISSN: 1743-422X
DOI
https://doi.org/10.1186/1743-422X-8-203

Other articles of this Issue 1/2011

Virology Journal 1/2011 Go to the issue