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RNA interference techniques as an approach to elucidating ranavirus gene function.
Author(s) -
Sample Robert Cressten,
Whitley Dexter,
Henegar Jeffrey,
Sullivan Donna C,
Chinchar Greg
Publication year - 2009
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.23.1_supplement.439.6
Subject(s) - biology , ranavirus , viral replication , virology , gene , rna interference , viral pathogenesis , rna , virus , genetics
Viruses of the genus Ranavirus , family Iridoviridae , are a group of large, icosahedral dsDNA viruses which have emerged as new pathogens of poikilothermic vertebrates. To confront their spread, it is vital to understand the mechanisms which control virus replication, host‐range, and pathogenesis. Therefore, Frog virus 3 (FV3), the ranavirus type species, was chosen as the model system. The FV3 genome has been completely sequenced and ~25% of the genes have recognized functions whereas the remaining ~75% have no known function. An antisense approach using RNA interference (RNAi) and antisense morpholinos (asMOs) was chosen to identify gene function and determine the requirements for replication in vitro and pathogenesis. The major capsid protein (MCP), the viral homologue of RNA Polymerase II (vPol‐IIα}), myristolated membrane protein (MMP), ICP‐46 (46K), and the cytosine DNA‐methyltransferase (DMT) were targeted using gene specific siRNAs or asMOs. Reductions of >80% in gene expression and >90% in viral titers were observed along with reductions in cytopathic effect. Transmission electron microscopy revealed reductions in virion assembly and production of atypical elements. These results support antisense techniques as a way of identifying and elucidating the function of genes involved in viral replication and pathogenesis.