Characterization of a full-length infectious cDNA clone and a GFP reporter derivative of the oncolytic picornavirus SVV-001

John T. Poirier, P. Seshidhar Reddy, Neeraja Idamakanti, Shawn S. Li, Kristine L. Stump, Kevin D. Burroughs, Paul L. Hallenbeck, Charles M. Rudin

Research output: Contribution to journalArticle

Abstract

Seneca Valley virus (SVV-001) is an oncolytic picornavirus with selective tropism for a subset of human cancers with neuroendocrine differentiation. To characterize further the specificity of SVV- 001 and its patterns and kinetics of intratumoral spread, bacterial plasmids encoding a cDNA clone of the full-length wild-type virus and a derivative virus expressing GFP were generated. The full-length cDNA of the SVV-001 RNA genome was cloned into a bacterial plasmid under the control of the T7 core promoter sequence to create an infectious cDNA clone, pNTX-09. A GFP reporter virus cDNA clone, pNTX-11, was then generated by cloning a fusion protein of GFP and the 2A protein from foot-and-mouth disease virus immediately following the native SVV-001 2A sequence. Recombinant GFP-expressing reporter virus, SVV-GFP, was rescued from cells transfected with in vitro RNA transcripts from pNTX-11 and propagated in cell culture. The proliferation kinetics of SVV-001 and SVV-GFP were indistinguishable. The SVV-GFP reporter virus was used to determine that a subpopulation of permissive cells is present in small-cell lung cancer cell lines previously thought to lack permissivity to SVV-001. Finally, it was shown that SVV-GFP administered to tumour-bearing animals homes in to and infects tumours whilst having no detectable tropism for normal mouse tissues at 1×1011 viral particles kg-1, a dose equivalent to that administered in ongoing clinical trials. These infectious clones will be of substantial value in further characterizing the biology of this virus and as a backbone for the generation of additional oncolytic derivatives.

Original languageEnglish (US)
Pages (from-to)2606-2613
Number of pages8
JournalJournal of General Virology
Volume93
Issue numberPART 12
DOIs
StatePublished - Dec 1 2012

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Picornaviridae
Complementary DNA
Clone Cells
Viruses
Tropism
Plasmids
RNA
Neoplasms
Small Cell Lung Carcinoma
Virion
Organism Cloning
Cell Culture Techniques
Clinical Trials
Genome
Cell Line

ASJC Scopus subject areas

  • Virology

Cite this

Poirier, J. T., Seshidhar Reddy, P., Idamakanti, N., Li, S. S., Stump, K. L., Burroughs, K. D., ... Rudin, C. M. (2012). Characterization of a full-length infectious cDNA clone and a GFP reporter derivative of the oncolytic picornavirus SVV-001. Journal of General Virology, 93(PART 12), 2606-2613. https://doi.org/10.1099/vir.0.046011-0

Characterization of a full-length infectious cDNA clone and a GFP reporter derivative of the oncolytic picornavirus SVV-001. / Poirier, John T.; Seshidhar Reddy, P.; Idamakanti, Neeraja; Li, Shawn S.; Stump, Kristine L.; Burroughs, Kevin D.; Hallenbeck, Paul L.; Rudin, Charles M.

In: Journal of General Virology, Vol. 93, No. PART 12, 01.12.2012, p. 2606-2613.

Research output: Contribution to journalArticle

Poirier, JT, Seshidhar Reddy, P, Idamakanti, N, Li, SS, Stump, KL, Burroughs, KD, Hallenbeck, PL & Rudin, CM 2012, 'Characterization of a full-length infectious cDNA clone and a GFP reporter derivative of the oncolytic picornavirus SVV-001', Journal of General Virology, vol. 93, no. PART 12, pp. 2606-2613. https://doi.org/10.1099/vir.0.046011-0
Poirier, John T. ; Seshidhar Reddy, P. ; Idamakanti, Neeraja ; Li, Shawn S. ; Stump, Kristine L. ; Burroughs, Kevin D. ; Hallenbeck, Paul L. ; Rudin, Charles M. / Characterization of a full-length infectious cDNA clone and a GFP reporter derivative of the oncolytic picornavirus SVV-001. In: Journal of General Virology. 2012 ; Vol. 93, No. PART 12. pp. 2606-2613.
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