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Herpes Simplex Virus Mutant Generation and Dual-Detection Methods for Gaining Insight into Latent/Lytic Cycles In Vivo

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Herpes Simplex Virus

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1144))

Abstract

Two important components to a useful strategy to examine viral gene regulation in vivo are (1) a highly efficient protocol to generate viral mutants that limits undesired mutation and retains full replication competency in vivo and (2) an efficient system to detect and quantify viral promoter activity in rare cells in vivo. Our strategy and protocols for generating, characterizing, and employing HSV viral promoter/reporter mutants in vivo are provided in this two-part chapter.

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References

  1. Whitley RJ (2001) Chapter 73: Herpes simplex viruses. In: Knipe DM, Howley PM, Griffin DE, Lamb RA, Martin MA (eds) Field’s virology. Lippincott Williams & Wilkins, Philadelphia, pp 2461–2510

    Google Scholar 

  2. Whitley RJ (2002) Herpes simplex virus infection. Semin Pediatr Infect Dis 13:6–11

    Article  PubMed  Google Scholar 

  3. Wagner EK, Bloom DC (1997) Experimental investigation of herpes simplex virus latency. Clin Microbiol Rev 10:419–443

    PubMed Central  PubMed  CAS  Google Scholar 

  4. Sawtell NM (1998) The probability of in vivo reactivation of herpes simplex virus type 1 increases with the number of latently infected neurons in the ganglia. J Virol 72:6888–6892

    PubMed Central  PubMed  CAS  Google Scholar 

  5. Sawtell NM, Thompson RL (1992) Rapid in vivo reactivation of herpes simplex virus in latently infected murine ganglionic neurons after transient hyperthermia. J Virol 66:2150–2156

    PubMed Central  PubMed  CAS  Google Scholar 

  6. Rock DL, Nesburn AB, Ghiasi H et al (1987) Detection of latency-related viral RNAs in trigeminal ganglia of rabbits latently infected with herpes simplex virus type 1. J Virol 61: 3820–3826

    PubMed Central  PubMed  CAS  Google Scholar 

  7. Wechsler SL, Nesburn AB, Watson R et al (1988) Fine mapping of the latency-related gene of herpes simplex virus type 1: alternative splicing produces distinct latency-related RNAs containing open reading frames. J Virol 62: 4051–4058

    PubMed Central  PubMed  CAS  Google Scholar 

  8. Shimeld C, Hill T, Blyth B, Easty D (1989) An improved model of recurrent herpetic eye disease in mice. Curr Eye Res 8:1193–1205

    Article  PubMed  CAS  Google Scholar 

  9. LeBlanc RA, Pesnicak L, Godleski M, Straus SE (1999) Treatment of HSV-1 infection with immunoglobulin or acyclovir: comparison of their effects on viral spread, latency, and reactivation. Virology 262:230–236

    Article  PubMed  CAS  Google Scholar 

  10. Gierasch WW, Zimmerman DL, Ward SL et al (2006) Construction and characterization of bacterial artificial chromosomes containing HSV-1 strains 17 and KOS. J Virol Methods 135:197–206

    Article  PubMed  CAS  Google Scholar 

  11. Du T, Zhou G, Khan S et al (2010) Disruption of HDAC/CoREST/REST repressor by dnREST reduces genome silencing and increases virulence of herpes simplex virus. Proc Natl Acad Sci U S A 107:15904–15909

    Article  PubMed Central  PubMed  Google Scholar 

  12. Sawtell NM (2003) Quantitative analysis of herpes simplex virus reactivation in vivo demonstrates that reactivation in the nervous system is not inhibited at early times postinoculation. J Virol 77:4127–4138

    Article  PubMed Central  PubMed  CAS  Google Scholar 

  13. Sawtell NM (2005) Detection and quantification of the rare latently infected cell undergoing herpes simplex virus transcriptional activation in the nervous system in vivo. Methods Mol Biol 292:57–72

    PubMed  CAS  Google Scholar 

  14. Thompson RL, Preston CM, Sawtell NM (2009) De novo synthesis of VP16 coordinates the exit from HSV latency in vivo. PLoS Pathog 5:e1000352

    Article  PubMed Central  PubMed  CAS  Google Scholar 

  15. Thompson RL, Sawtell NM (2006) Evidence that the herpes simplex virus type 1 ICP0 protein does not initiate reactivation from latency in vivo. J Virol 80:10919–10930

    Article  PubMed Central  PubMed  CAS  Google Scholar 

  16. Thompson RL, Shieh MT, Sawtell NM (2003) Analysis of herpes simplex virus ICP0 promoter function in sensory neurons during acute infection, establishment of latency, and reactivation in vivo. J Virol 77:12319–12330

    Article  PubMed Central  PubMed  CAS  Google Scholar 

  17. Sawtell NM (1997) Comprehensive quantification of herpes simplex virus latency at the single-cell level. J Virol 71:5423–5431

    PubMed Central  PubMed  CAS  Google Scholar 

  18. Thompson RL, Sawtell NM (2000) Replication of herpes simplex virus type 1 within trigeminal ganglia is required for high frequency but not high viral genome copy number latency. J Virol 74:965–974

    Article  PubMed Central  PubMed  CAS  Google Scholar 

  19. Thompson RL, Sawtell NM (2011) The herpes simplex virus type 1 latency associated transcript locus is required for the maintenance of reactivation competent latent infections. J Neurovirol 17:552–558

    Article  PubMed Central  PubMed  CAS  Google Scholar 

  20. Helander KG (2000) Formaldehyde prepared from paraformaldehyde is stable. Biotech Histochem 75:19–22

    Article  PubMed  CAS  Google Scholar 

  21. Cunningham C, Davison AJ (1993) A cosmid-based system for constructing mutants of herpes simplex virus type 1. Virology 197:116–124

    Article  PubMed  CAS  Google Scholar 

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Author information

Authors and Affiliations

  1. Division of Infectious Diseases, Department of Pediatrics, Cincinnati Children’s Hospital Medical Center, 3333 Burnet Ave., MLC 7017, Cincinnati, OH, 45229-3039, USA

    Nancy M. Sawtell Ph.D.

  2. Department of Molecular Genetics, Biochemistry, and Microbiology, University of Cincinnati College of Medicine, Cincinnati, OH, 45267-0524, USA

    Richard L. Thompson

Authors
  1. Nancy M. Sawtell Ph.D.
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  2. Richard L. Thompson
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Corresponding author

Correspondence to Nancy M. Sawtell Ph.D. .

Editor information

Editors and Affiliations

  1. Centre for Virus Research, Westmead Millennium Institute, Westmead, New South Wales, Australia

    Russell J. Diefenbach

  2. Institute of Virology, University of Zürich, Zürich, Switzerland

    Cornel Fraefel

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Sawtell, N.M., Thompson, R.L. (2014). Herpes Simplex Virus Mutant Generation and Dual-Detection Methods for Gaining Insight into Latent/Lytic Cycles In Vivo. In: Diefenbach, R., Fraefel, C. (eds) Herpes Simplex Virus. Methods in Molecular Biology, vol 1144. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-0428-0_9

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  • DOI: https://doi.org/10.1007/978-1-4939-0428-0_9

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-0427-3

  • Online ISBN: 978-1-4939-0428-0

  • eBook Packages: Springer Protocols

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