Firefly Luciferase as a Marker for Herpesvirus (pseudorabies virus) Replication in Vitro and in Vivo

Sz, Ferenc Kovfics; Mettenleiter, Thomas C.

doi:10.1099/0022-1317-72-12-2999

Cited by 28 publications

(1 citation statement)

References 33 publications

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“…In some previous studies, the firefly luciferase expression cassette was recombined into PRV for expression [ 55 , 56 ], but few studies have reported imaging of PRV in vivo. In this study, the imaging characteristics were analysed in vivo in mice infected with different doses of the recombinant virus rPRV HN1201-EGFP-Luc.…”

Section: Discussionmentioning

confidence: 99%

CRISPR/Cas9-based generation of a recombinant double-reporter pseudorabies virus and its characterization in vitro and in vivo

Cheng

et al. 2021

Vet Res

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Pseudorabies, caused by pseudorabies virus (PRV) variants, has broken out among commercial PRV vaccine-immunized swine herds and resulted in major economic losses to the pig industry in China since late 2011. However, the mechanism of virulence enhancement of variant PRV is currently unclear. Here, a recombinant PRV (rPRV HN1201-EGFP-Luc) with stable expression of enhanced green fluorescent protein (EGFP) and firefly luciferase as a double reporter virus was constructed on the basis of the PRV variant HN1201 through CRISPR/Cas9 gene-editing technology coupled with two sgRNAs. The biological characteristics of the recombinant virus and its lethality to mice were similar to those of the parental strain and displayed a stable viral titre and luciferase activity through 20 passages. Moreover, bioluminescence signals were detected in mice at 12 h after rPRV HN1201-EGFP-Luc infection. Using the double reporter PRV, we also found that 25-hydroxycholesterol had a significant inhibitory effect on PRV both in vivo and in vitro. These results suggested that the double reporter PRV based on PRV variant HN1201 should be an excellent tool for basic virology studies and evaluating antiviral agents.

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Section: Discussionmentioning

confidence: 99%