RNAi-dependent and -independent antiviral phenotypes of chromosomally integrated shRNA clones: role of VASP in respiratory syncytial virus growth.
Stable RNA interference (RNAi) is commonly achieved by recombinant expression of short hairpin RNA (shRNA). To generate virus-resistant cell lines, we cloned a shRNA cassette against the phosphoprotein gene of respiratory syncytial virus (RSV) into a polIII-driven plasmid vector. Analysis of individual stable transfectants showed a spectrum of RSV resistance correlating with the levels of shRNA expressed from different chromosomal locations. Interestingly, resistance in a minority of clones was due to mono-allelic disruption of the cellular gene for vasodilator-stimulated phosphoprotein (VASP). Thus, pure clones of chromosomally integrated DNA-directed RNAi can exhibit gene disruption phenotypes resembling but unrelated to RNAi.
Musiyenko
Alla
A
Department of Biochemistry and Molecular Biology, College of Medicine, University of South Alabama, 307 University Blvd., Mobile, AL 36688-0002, USA.
Bitko
Vira
V
Barik
Sailen
S
D001483:Base Sequence; D015815:Cell Adhesion Molecules / Q000235:genetics / Q000502:physiology*; D002877:Chromosomes, Human; D003001:Cloning, Molecular / Q000379:methods; D005822:Genetic Vectors; D006367:HeLa Cells; D006801:Humans; D007109:Immunity; D008840:Microfilament Proteins / Q000235:genetics / Q000502:physiology*; D009838:Oligodeoxyribonucleotides / Q000138:chemical synthesis / Q000235:genetics* / Q000494:pharmacology; D010750:Phosphoproteins / Q000235:genetics / Q000502:physiology*; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000502:physiology*; D012136:Respiratory Syncytial Viruses / Q000254:growth & development / Q000276:immunology*; D014162:Transfection; D000098662:Vasodilator-Stimulated Phosphoprotein
D015815:Cell Adhesion Molecules; D008840:Microfilament Proteins; D009838:Oligodeoxyribonucleotides; D010750:Phosphoproteins; D034741:RNA, Small Interfering; D000098662:Vasodilator-Stimulated Phosphoprotein