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European Journal of Clinical Microbiology & Infectious Diseases

Published in:

01-08-2008 | Article

siRNA silencing of angiotensin-converting enzyme 2 reduced severe acute respiratory syndrome-associated coronavirus replications in Vero E6 cells

Authors: C.-Y. Lu, H.-Y. Huang, T.-H. Yang, L.-Y. Chang, C.-Y. Lee, L.-M. Huang

Published in: European Journal of Clinical Microbiology & Infectious Diseases | Issue 8/2008

Abstract

The outbreak of severe acute respiratory syndrome (SARS) in 2002–2003 has had a significant impact worldwide. No effective prophylaxis or treatment for SARS is available up to now. Angiotensin-converting enzyme 2 (ACE2) is the cellular receptor for SARS-associated coronavirus (SARS-CoV). By expressing a U6 promoter-driven small interfering RNA containing sequences homologous to part of ACE2 mRNA, we successfully silenced ACE2 expression in Vero E6 cells. By detecting negative strand SARS-CoV RNA and measuring RNA copy numbers of SARS-CoV by real-time reverse transcription polymerase chain reaction (RT-PCR), we demonstrated that SARS-CoV infection was reduced in the ACE2-silenced cell lines. These findings support the involvement of ACE2 in SARS-CoV infections and provide a basis for further studies on potential use of siRNA targeting ACE2 as a preventive or therapeutic strategy for SARS.

Groneberg DA, Poutanen SM, Low DE, Lode H, Welte T, Zabel P (2005) Treatment and vaccines for severe acute respiratory syndrome. Lancet Infect Dis 5:147–155PubMed

Hsueh PR, Hsiao CH, Yeh SH et al (2003) Microbiologic characteristics, serologic responses, and clinical manifestations in severe acute respiratory syndrome, Taiwan. Emerg Infect Dis 9:1163–1167PubMed

Gallagher TM, Buchmeier MJ (2001) Coronavirus spike proteins in viral entry and pathogenesis. Virology 279:371–374PubMedCrossRef

Li W, Moore MJ, Vasilieva N et al (2003) Angiotensin-converting enzyme 2 is a functional receptor for the SARS coronavirus. Nature 426:450–454PubMedCrossRef

Donoghue M, Hsieh F, Baronas E et al (2000) A novel angiotensin-converting enzyme-related carboxypeptidase (ACE2) converts angiotensin I to angiotensin 1–9. Circ Res 87:E1–E9PubMed

Harmer D, Gilbert M, Borman R, Clark KL (2002) Quantitative mRNA expression profiling of ACE 2, a novel homologue of angiotensin converting enzyme. FEBS Lett 532:107–110PubMedCrossRef

Agrawal N, Dasaradhi PV, Mohmmed A, Malhotra P, Bhatnagar RK, Mukherjee SK (2003) RNA interference: biology, mechanism, and applications. Microbiol Mol Biol Rev 67:657–685PubMedCrossRef

Fire A, Xu S, Montgomery MK, Kostas SA, Driver SE, Mello CC (1998) Potent and specific genetic interference by double-stranded RNA in Caenorhabditis elegans. Nature 391:806–811PubMedCrossRef

Voinnet O (2005) Induction and suppression of RNA silencing: insights from viral infections. Nat Rev Genet 6:206–220PubMedCrossRef

10.

Ksiazek TG, Erdman D, Goldsmith CS et al (2003) A novel coronavirus associated with severe acute respiratory syndrome. N Engl J Med 348:1953–1966PubMedCrossRef

11.

Drosten C, Gunther S, Preiser W et al (2003) Identification of a novel coronavirus in patients with severe acute respiratory syndrome. N Engl J Med 348:1967–1976PubMedCrossRef

12.

Li T, Zhang Y, Fu L et al (2005) siRNA targeting the leader sequence of SARS-CoV inhibits virus replication. Gene Ther 12:751–761PubMedCrossRef

13.

Qin ZL, Zhao P, Zhang XL et al (2004) Silencing of SARS-CoV spike gene by small interfering RNA in HEK 293T cells. Biochem Biophys Res Commun 324:1186–1193PubMedCrossRef

14.

Zhang Y, Li T, Fu L et al (2004) Silencing SARS-CoV Spike protein expression in cultured cells by RNA interference. FEBS Lett 560:141–146PubMedCrossRef

15.

Shi Y, Yang DH, Xiong J, Jia J, Huang B, Jin YX (2005) Inhibition of genes expression of SARS coronavirus by synthetic small interfering RNAs. Cell Res 15:193–200PubMedCrossRef

16.

Qin ZL, Zhao P, Cao MM, Qi ZT (2007) siRNAs targeting terminal sequences of the SARS-associated coronavirus membrane gene inhibit M protein expression through degradation of M mRNA. J Virol Methods 145:146–154PubMedCrossRef

17.

Wang Z, Ren L, Zhao X et al (2004) Inhibition of severe acute respiratory syndrome virus replication by small interfering RNAs in mammalian cells. J Virol 78:7523–7527PubMedCrossRef

18.

Ni B, Shi X, Li Y, Gao W, Wang X, Wu Y (2005) Inhibition of replication and infection of severe acute respiratory syndrome-associated coronavirus with plasmid-mediated interference RNA. Antivir Ther 10:527–533PubMed

19.

Akerstrom S, Mirazimi A, Tan YJ (2007) Inhibition of SARS-CoV replication cycle by small interference RNAs silencing specific SARS proteins, 7a/7b, 3a/3b and S. Antiviral Res 73:219–227PubMedCrossRef

20.

Wu CJ, Huang HW, Liu CY, Hong CF, Chan YL (2005) Inhibition of SARS-CoV replication by siRNA. Antiviral Res 65:45–48PubMedCrossRef

21.

Li BJ, Tang Q, Cheng D et al (2005) Using siRNA in prophylactic and therapeutic regimens against SARS coronavirus in Rhesus macaque. Nat Med 11:944–951PubMed

22.

Wu CJ, Chan YL (2006) Antiviral applications of RNAi for coronavirus. Expert Opin Investig Drugs 15:89–97PubMedCrossRef

23.

Chang Z, Babiuk LA, Hu J (2007) Therapeutic and prophylactic potential of small interfering RNAs against severe acute respiratory syndrome: progress to date. BioDrugs 21:9–15PubMedCrossRef

24.

Anderson J, Akkina R (2005) CXCR4 and CCR5 shRNA transgenic CD34+ cell derived macrophages are functionally normal and resist HIV-1 infection. Retrovirology 2:53PubMedCrossRef

25.

Novina CD, Murray MF, Dykxhoorn DM et al (2002) siRNA-directed inhibition of HIV-1 infection. Nat Med 8:681–686PubMed

26.

Hattermann K, Muller MA, Nitsche A, Wendt S, Donoso Mantke O, Niedrig M (2005) Susceptibility of different eukaryotic cell lines to SARS-coronavirus. Arch Virol 150:1023–1031PubMedCrossRef

27.

Hofmann H, Geier M, Marzi A et al (2004) Susceptibility to SARS coronavirus S protein-driven infection correlates with expression of angiotensin converting enzyme 2 and infection can be blocked by soluble receptor. Biochem Biophys Res Commun 319:1216–1221PubMedCrossRef

28.

Nie Y, Wang P, Shi X et al (2004) Highly infectious SARS-CoV pseudotyped virus reveals the cell tropism and its correlation with receptor expression. Biochem Biophys Res Commun 321:994–1000PubMedCrossRef

29.

Mossel EC, Huang C, Narayanan K, Makino S, Tesh RB, Peters CJ (2005) Exogenous ACE2 expression allows refractory cell lines to support severe acute respiratory syndrome coronavirus replication. J Virol 79:3846–3850PubMedCrossRef

30.

Jeffers SA, Tusell SM, Gillim-Ross L et al (2004) CD209L (L-SIGN) is a receptor for severe acute respiratory syndrome coronavirus. Proc Natl Acad Sci USA 101:15748–15753PubMedCrossRef

31.

He ML, Zheng BJ, Chen Y et al (2006) Kinetics and synergistic effects of siRNAs targeting structural and replicase genes of SARS-associated coronavirus. FEBS Lett 580:2414–2420PubMedCrossRef

32.

de Lang A, Osterhaus AD, Haagmans BL (2006) Interferon-gamma and interleukin-4 downregulate expression of the SARS coronavirus receptor ACE2 in Vero E6 cells. Virology 353:474–481PubMedCrossRef

33.

Goulter AB, Goddard MJ, Allen JC, Clark KL (2004) ACE2 gene expression is up-regulated in the human failing heart. BMC Med 2:19PubMedCrossRef

34.

Crackower MA, Sarao R, Oudit GY et al (2002) Angiotensin-converting enzyme 2 is an essential regulator of heart function. Nature 417:822–828PubMedCrossRef

Title: siRNA silencing of angiotensin-converting enzyme 2 reduced severe acute respiratory syndrome-associated coronavirus replications in Vero E6 cells
Authors: C.-Y. Lu
H.-Y. Huang
T.-H. Yang
L.-Y. Chang
C.-Y. Lee
L.-M. Huang
Publication date: 01-08-2008
Publisher: Springer-Verlag
Published in: European Journal of Clinical Microbiology & Infectious Diseases / Issue 8/2008
Print ISSN: 0934-9723
Electronic ISSN: 1435-4373
DOI: https://doi.org/10.1007/s10096-008-0495-5

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