JC virus-iLOV fluorescent strains enable the detection of early and late viral protein expression
| Autor: | Igor J. Koralnik, Spyridon Chalkias, Xin Dang |
|---|---|
| Rok vydání: | 2015 |
| Předmět: |
Viral protein
viruses Drug Evaluation Preclinical JC virus Biology medicine.disease_cause Antiviral Agents Article Cell Line Viral Proteins Genes Reporter Virology medicine Humans Gene Recombination Genetic Multiple sclerosis Progressive multifocal leukoencephalopathy virus diseases medicine.disease JC Virus In vitro nervous system diseases Luminescent Proteins nervous system Cell culture Viral load |
| Zdroj: | Journal of Virological Methods. 223:25-29 |
| ISSN: | 0166-0934 |
| DOI: | 10.1016/j.jviromet.2015.07.006 |
| Popis: | JC virus (JCV) is highly prevalent in humans, and may cause progressive multifocal leukoencephalopathy (PML), JCV granule cell neuronopathy (JCV GCN), JCV encephalopathy (JCVE) and JCV meningitis (JCVM) in immunocompromised individuals. There is no treatment for JCV, and a growing number of multiple sclerosis patients treated with immunomodulatory medications have developed PML. Antiviral agents against JCV are therefore highly desirable but remain elusive, due to the difficulty of determining their effect in vitro. A JCV strain carrying a fluorescent protein gene would greatly simplify and accelerate the drug screening process. To achieve this goal, we selected the 366 bp improved Light, Oxygen or Voltage-sensing domain (iLOV) of plant phototropin gene and created two full-length JCV-iLOV constructs on the prototype JCV Mad1 backbone. The iLOV gene was inserted either before the early regulatory T gene (iLOV-T), or after the late Agno gene (iLOV-Agno). Both JCV iLOV strains were replication-competent in vitro and emitted a fluorescent signal detectable by confocal microscope, but JCV iLOV-T exhibited higher cellular and supernatant viral loads compared to JCV iLOV-Agno. JCV iLOV-T could also produce infectious pseudovirions. These data suggest that JCV iLOV constructs may become valuable tools for anti-JCV drug screening. |
| Databáze: | OpenAIRE |
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