Single-stranded positive-sense RNA viruses generated in days using infectious subgenomic amplicons Open Access

Abstract

Reverse genetics is a key methodology for producing genetically modified RNA viruses and deciphering cellular and viral biological properties, but methods based on the preparation of plasmid-based complete viral genomes are laborious and unpredictable. Here, both wild-type and genetically modified infectious RNA viruses were generated in days using the newly described ISA (infectious-subgenomic-amplicons) method. This new versatile and simple procedure may enhance our capacity to obtain infectious RNA viruses from PCR-amplified genetic material.

Loading

Article metrics loading...

/content/journal/jgv/10.1099/vir.0.068023-0
2014-11-01
2024-03-29
Loading full text...

Full text loading...

/deliver/fulltext/jgv/95/11/2462.html?itemId=/content/journal/jgv/10.1099/vir.0.068023-0&mimeType=html&fmt=ahah

References

  1. Aubry F., Vongsouvath M., Nougairède A., Phetsouvanh R., Sibounheuang B., Charrel R., Rattanavong S., Phommasone K., Sengvilaipraserth O.other authors 2013; Complete genome of a genotype I Japanese encephalitis virus isolated from a patient with encephalitis in Vientiane, Lao PDR. Genome Announc 1:e00157-12 [View Article][PubMed]
    [Google Scholar]
  2. Bacher J. M., Reiss B. D., Ellington A. D. 2002; Anticipatory evolution and DNA shuffling. Genome Biol 3:reviews1021.1–reviews1021.4 [View Article][PubMed]
    [Google Scholar]
  3. Bredenbeek P. J., Kooi E. A., Lindenbach B., Huijkman N., Rice C. M., Spaan W. J. 2003; A stable full-length yellow fever virus cDNA clone and the role of conserved RNA elements in flavivirus replication. J Gen Virol 84:1261–1268 [View Article][PubMed]
    [Google Scholar]
  4. Csörgo B., Fehér T., Tímár E., Blattner F. R., Pósfai G. 2012; Low-mutation-rate, reduced-genome Escherichia coli: an improved host for faithful maintenance of engineered genetic constructs. Microb Cell Fact 11:11 [View Article][PubMed]
    [Google Scholar]
  5. Edmonds J., van Grinsven E., Prow N., Bosco-Lauth A., Brault A. C., Bowen R. A., Hall R. A., Khromykh A. A. 2013; A novel bacterium-free method for generation of flavivirus infectious DNA by circular polymerase extension reaction allows accurate recapitulation of viral heterogeneity. J Virol 87:2367–2372 [View Article][PubMed]
    [Google Scholar]
  6. Gorchakov R., Wang E., Leal G., Forrester N. L., Plante K., Rossi S. L., Partidos C. D., Adams A. P., Seymour R. L.other authors 2012; Attenuation of Chikungunya virus vaccine strain 181/clone 25 is determined by two amino acid substitutions in the E2 envelope glycoprotein. J Virol 86:6084–6096 [View Article][PubMed]
    [Google Scholar]
  7. Gritsun T. S., Gould E. A. 1995; Infectious transcripts of tick-borne encephalitis virus, generated in days by RT-PCR. Virology 214:611–618 [View Article][PubMed]
    [Google Scholar]
  8. Gritsun T. S., Gould E. A. 1998; Development and analysis of a tick-borne encephalitis virus infectious clone using a novel and rapid strategy. J Virol Methods 76:109–120 [View Article][PubMed]
    [Google Scholar]
  9. Henchal E. A., Gentry M. K., McCown J. M., Brandt W. E. 1982; Dengue virus-specific and flavivirus group determinants identified with monoclonal antibodies by indirect immunofluorescence. Am J Trop Med Hyg 31:830–836[PubMed]
    [Google Scholar]
  10. Marillonnet S., Giritch A., Gils M., Kandzia R., Klimyuk V., Gleba Y. 2004; In planta engineering of viral RNA replicons: efficient assembly by recombination of DNA modules delivered by Agrobacterium. Proc Natl Acad Sci U S A 101:6852–6857 [View Article][PubMed]
    [Google Scholar]
  11. Mishin V. P., Cominelli F., Yamshchikov V. F. 2001; A ‘minimal’ approach in design of flavivirus infectious DNA. Virus Res 81:113–123 [View Article][PubMed]
    [Google Scholar]
  12. Polo S., Ketner G., Levis R., Falgout B. 1997; Infectious RNA transcripts from full-length dengue virus type 2 cDNA clones made in yeast. J Virol 71:5366–5374[PubMed]
    [Google Scholar]
  13. Pu S. Y., Wu R. H., Yang C. C., Jao T. M., Tsai M. H., Wang J. C., Lin H. M., Chao Y. S., Yueh A. 2011; Successful propagation of flavivirus infectious cDNAs by a novel method to reduce the cryptic bacterial promoter activity of virus genomes. J Virol 85:2927–2941 [View Article][PubMed]
    [Google Scholar]
  14. Pu S. Y., Wu R. H., Tsai M. H., Yang C. C., Chang C. M., Yueh A. 2014; A novel approach to propagate flavivirus infectious cDNA clones in bacteria by introducing tandem repeat sequences upstream of virus genome. J Gen Virol 95:1493–1503 [View Article][PubMed]
    [Google Scholar]
  15. Reed L. J., Muench H. 1938; A simple method of estimating fifty per cent endpoints. Am J Hyg 27:493–497
    [Google Scholar]
  16. Rothberg J. M., Hinz W., Rearick T. M., Schultz J., Mileski W., Davey M., Leamon J. H., Johnson K., Milgrew M. J.other authors 2011; An integrated semiconductor device enabling non-optical genome sequencing. Nature 475:348–352 [View Article][PubMed]
    [Google Scholar]
  17. Ruggli N., Rice C. M. 1999; Functional cDNA clones of the Flaviviridae: strategies and applications. Adv Virus Res 53:183–207 [View Article][PubMed]
    [Google Scholar]
  18. Schoggins J. W., Dorner M., Feulner M., Imanaka N., Murphy M. Y., Ploss A., Rice C. M. 2012; Dengue reporter viruses reveal viral dynamics in interferon receptor-deficient mice and sensitivity to interferon effectors in vitro. Proc Natl Acad Sci U S A 109:14610–14615 [View Article][PubMed]
    [Google Scholar]
  19. Siridechadilok B., Gomutsukhavadee M., Sawaengpol T., Sangiambut S., Puttikhunt C., Chin-inmanu K., Suriyaphol P., Malasit P., Screaton G., Mongkolsapaya J. 2013; A simplified positive-sense-RNA virus construction approach that enhances analysis throughput. J Virol 87:12667–12674 [View Article][PubMed]
    [Google Scholar]
  20. Stang A., Korn K., Wildner O., Uberla K. 2005; Characterization of virus isolates by particle-associated nucleic acid PCR. J Clin Microbiol 43:716–720 [View Article][PubMed]
    [Google Scholar]
  21. Suzuki R., de Borba L., Duarte dos Santos C. N., Mason P. W. 2007; Construction of an infectious cDNA clone for a Brazilian prototype strain of dengue virus type 1: characterization of a temperature-sensitive mutation in NS1. Virology 362:374–383 [View Article][PubMed]
    [Google Scholar]
  22. Zhang F., Huang Q., Ma W., Jiang S., Fan Y., Zhang H. 2001; Amplification and cloning of the full-length genome of Japanese encephalitis virus by a novel long RT-PCR protocol in a cosmid vector. J Virol Methods 96:171–182 [View Article][PubMed]
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jgv/10.1099/vir.0.068023-0
Loading
/content/journal/jgv/10.1099/vir.0.068023-0
Loading

Data & Media loading...

Supplements

Supplementary material 1

PDF

Most cited Most Cited RSS feed