Examination of antisense RNA and oligodeoxynucleotides as potential inhibitors of avian leukosis virus replication in RP30 cells.
Avian leukosis virus (ALV) is an economically important pathogen of chickens. Both antisense RNA and antisense oligodeoxynucleotides (ODN) have been used to diminish the replication and spread of other retroviruses. The use of antisense RNA and ODN to inhibit ALV replication has been examined in cultured RP30 cells. Using an expression system that constitutively transcribes antisense ALV RNA, one transfected cell clone showed a significant reduction in virus growth. However, this effect was not reproducibly observed in other transfected cell lines or in cells in which the antisense transcript was expressed from a regulatable promoter, even though a substantial amount of antisense transcript was generated. Antisense ODN complementary to several different target sites near the 5' end of the ALV genome were also tested for antiviral activity, by comparison of antisense ODN effects to those of randomized sequence controls. An antisense ODN complementary to the ALV primer binding site demonstrated a reproducible reduction in viral replication. However, when the corresponding region was specifically employed as a target for intracellular antisense RNA expression, there again was no significant inhibition of ALV. These results suggest that in vivo expression of antisense RNA is unlikely to be an effective way to generate transgenic poultry that are resistant to field strains of ALV.
Kim
K E
KE
Department of Microbiology, Michigan State University, East Lansing 48824, USA.
Salter
D W
DW
Dodgson
J B
JB
D000818:Animals; D001354:Avian Leukosis Virus / Q000235:genetics / Q000502:physiology*; D002461:Cell Line, Transformed; D015870:Gene Expression / Q000187:drug effects; D005814:Genes, Viral; D008214:Lymphocytes / Q000821:virology*; D020319:Oligodeoxyribonucleotides, Antisense / Q000494:pharmacology*; D011201:Poultry Diseases / Q000821:virology; D016372:RNA, Antisense / Q000235:genetics / Q000494:pharmacology*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013752:Tetracycline / Q000494:pharmacology; D014162:Transfection; D014422:Turkeys; D014779:Virus Replication / Q000187:drug effects*
D020319:Oligodeoxyribonucleotides, Antisense; D016372:RNA, Antisense; D013752:Tetracycline