Abstract
RNA interference is an important tool for gene silencing. However, its application to primary cultured cells has been limited by low transfection efficiencies. In this work we developed a vector which encodes both siRNA and red fluorescent protein. Using this vector we could markedly suppress green fluorescent protein (GFP) and bim an endogenous gene. Primary cultured cortical neurons transfected with siRNA against doublecortin showed that doublecortin expression was significantly inhibited in nearly all the transfected neurons. This vector identifies the transfected cells and should be useful for loss-of-gene function studies in neurons.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Animals
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Apoptosis Regulatory Proteins
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Bcl-2-Like Protein 11
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Carrier Proteins / genetics
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Carrier Proteins / metabolism
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Cells, Cultured
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Doublecortin Domain Proteins
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Genes, Reporter*
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Genetic Vectors* / genetics
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Genetic Vectors* / metabolism
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Green Fluorescent Proteins / genetics
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Green Fluorescent Proteins / metabolism*
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Humans
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Membrane Proteins / genetics
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Membrane Proteins / metabolism
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Microtubule-Associated Proteins / genetics
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Microtubule-Associated Proteins / metabolism
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Neurons / cytology
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Neurons / physiology*
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Neuropeptides / genetics
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Neuropeptides / metabolism
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Proto-Oncogene Proteins / genetics
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Proto-Oncogene Proteins / metabolism
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RNA Interference
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RNA, Small Interfering* / genetics
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RNA, Small Interfering* / metabolism
Substances
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Apoptosis Regulatory Proteins
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BCL2L11 protein, human
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Bcl-2-Like Protein 11
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Carrier Proteins
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Doublecortin Domain Proteins
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Membrane Proteins
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Microtubule-Associated Proteins
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Neuropeptides
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Proto-Oncogene Proteins
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RNA, Small Interfering
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Green Fluorescent Proteins