1887

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.

  • This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
Loading

Article metrics loading...

/content/journal/jgv/10.1099/vir.0.068023-0
2014-11-01
2022-05-29
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 this month Most Cited RSS feed

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error