In vitro inhibition of porcine hemagglutinating encephalomyelitis virus replication with siRNAs targeting the spike glycoprotein and replicase polyprotein genes.

Abstract:

OBJECTIVE:The specific effect of rna interference on the replication of porcine hemagglutinating encephalomyelitis virus (phe-cov) was explored. METHODS:Four species of small interfering RNA (siRNA), targeting different regions of the PHE-CoV spike glycoprotein and replicase polyprotein genes, were prepared by in vitro transcription. After transfection of PK-15 cells with each of the siRNAs followed by infection with PHE-CoV, the cytopathic effect (CPE) was examined by phase-contrast microscope, and viral proliferation within cells was examined by indirect immunofluorescence microscopy, hemagglutination (HA) test, TCID(50) assay and real-time RT-PCR. RESULTS:Examination of CPE demonstrated that the four siRNAs were capable of protecting cells against PHE-CoV invasion with very high specificity and efficiency. At 48 h post-infection, only a few siRNA-treated cells were positive for viral antigen staining, whereas most untreated virus-infected cells were positive. Transfection with siRNAs also suppressed the production of infectious virus by up to 18- to 32-fold as assessed by a HA test and 93- to 494-fold as assessed by TCID(50) assay. Furthermore, treatment with siRNAs caused a 53-91% reduction in the viral genome copy number as assessed by real-time RT-PCR. CONCLUSION:These results suggested that the four species of siRNAs can efficiently inhibit PHE-CoV genome replication and infectious virus production.

journal_name

Intervirology

journal_title

Intervirology

authors

Lan Y,Lu H,Zhao K,He W,Chen K,Wang G,Song D,Gao F

doi

10.1159/000323523

subject

Has Abstract

pub_date

2012-01-01 00:00:00

pages

53-61

issue

1

eissn

0300-5526

issn

1423-0100

pii

000323523

journal_volume

55

pub_type

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