Abstract
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.
Publication types
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Research Support, N.I.H., Extramural
MeSH terms
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Base Sequence
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Cell Adhesion Molecules / genetics
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Cell Adhesion Molecules / physiology*
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Chromosomes, Human
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Cloning, Molecular / methods
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Genetic Vectors
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HeLa Cells
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Humans
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Immunity*
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Microfilament Proteins / genetics
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Microfilament Proteins / physiology*
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Oligodeoxyribonucleotides / chemical synthesis
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Oligodeoxyribonucleotides / genetics*
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Oligodeoxyribonucleotides / pharmacology
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Phosphoproteins / genetics
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Phosphoproteins / physiology*
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RNA Interference*
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RNA, Small Interfering / physiology*
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Respiratory Syncytial Viruses / growth & development
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Respiratory Syncytial Viruses / immunology*
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Transfection
Substances
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Cell Adhesion Molecules
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Microfilament Proteins
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Oligodeoxyribonucleotides
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Phosphoproteins
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RNA, Small Interfering
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vasodilator-stimulated phosphoprotein