title: Antisense approaches for elucidating ranavirus gene function in an infected fish cell line.
abstract: Viral virulence/immune evasion strategies and host anti-viral responses represent different sides of the continuing struggle between virus and host survival. To identify virus-encoding molecules whose function is to subvert or blunt host immune responses, we have adapted anti-sense approaches to knock down the expression of specific viral gene products. Our intention is to correlate knock down with loss of function and thus infer the role of a given viral gene. As a starting point in this process we have targeted several structural and catalytic genes using antisense morpholino oligonucleotides (asMO) and small, interfering RNAs (siRNA). In proof of concept experiments we show the feasibility of this approach and describe recent work targeting five frog virus 3 genes. Our results indicate that both 46K and 32R, two immediate-early viral proteins, are essential for replication in vitro, and confirm earlier findings that the major capsid protein, the largest subunit of the viral homolog of RNA polymerase II, and the viral DNA methyltransferase are also essential for replication in cell culture.
authors.affiliation: Department of Microbiology, University of Mississippi Medical Center, Jackson, MS 39216, United States.
authors.forename: D S
authors.initials: DS
authors.lastname: Whitley
authors.forename: R C
authors.initials: RC
authors.lastname: Sample
authors.forename: A R
authors.initials: AR
authors.lastname: Sinning
authors.forename: J
authors.initials: J
authors.lastname: Henegar
authors.forename: V G
authors.initials: VG
authors.lastname: Chinchar
mesh_terms: D000818:Animals; D036022:Capsid Proteins / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D002460:Cell Line; D015254:DNA Modification Methylases / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D004266:DNA Virus Infections / Q000276:immunology / Q000821:virology*; D005399:Fishes; D055785:Gene Knockdown Techniques; D054884:Host-Pathogen Interactions; D057131:Immune Evasion; D016376:Oligonucleotides, Antisense / Q000235:genetics; D034741:RNA, Small Interfering / Q000235:genetics; D017995:Ranavirus / Q000472:pathogenicity / Q000502:physiology*; D014764:Viral Proteins / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D014779:Virus Replication
chemical_list: D036022:Capsid Proteins; D016376:Oligonucleotides, Antisense; D034741:RNA, Small Interfering; D014764:Viral Proteins; C081132:major capsid protein, Iridovirus; D015254:DNA Modification Methylases