Abstract
Drug-inducible systems allowing the control of gene expression in mammalian cells are invaluable tools for genetic research, and could also fulfill essential roles in gene- and cell-based therapy. Currently available systems, however, often have limited in vivo functionality because of leakiness, insufficient levels of induction, lack of tissue specificity or prohibitively complicated designs. Here we describe a lentiviral vector-based, conditional gene expression system for drug-controllable expression of polymerase (Pol) II promoter-driven transgenes or Pol III promoter-controlled sequences encoding small inhibitory hairpin RNAs (shRNAs). This system has great robustness and versatility, governing tightly controlled gene expression in cell lines, in embryonic or hematopoietic stem cells, in human tumors xenotransplanted into nude mice, in the brain of rats injected intraparenchymally with the vector, and in transgenic mice generated by infection of fertilized oocytes. These results open up promising perspectives for basic or translational research and for the development of gene-based therapeutics.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Animals
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Bacterial Proteins / genetics
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Brain / metabolism
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Carrier Proteins / genetics
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Cell Differentiation / genetics
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Cell Line
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Cell Line, Tumor
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DNA Polymerase II / genetics
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DNA Polymerase III / genetics
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Doxycycline / pharmacology
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Female
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GATA1 Transcription Factor / genetics
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Gene Expression Regulation / drug effects
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Gene Expression Regulation / genetics*
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Gene Silencing
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Gene Targeting / methods*
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Genetic Engineering / methods
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Genetic Vectors / genetics
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Glial Cell Line-Derived Neurotrophic Factor / genetics
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Glial Cell Line-Derived Neurotrophic Factor / 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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Lentivirus / genetics
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Male
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Mice
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Mice, Inbred Strains
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Mice, Nude
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Mice, Transgenic
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Promoter Regions, Genetic / genetics
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RNA Interference*
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RNA, Small Interfering / genetics
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Rats
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Rats, Sprague-Dawley
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Rats, Wistar
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Repressor Proteins / genetics
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Stem Cells / cytology
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Stem Cells / drug effects
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Stem Cells / metabolism
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Transfection
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Transgenes / genetics
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Tumor Suppressor Protein p53 / genetics
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Xenograft Model Antitumor Assays
Substances
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Bacterial Proteins
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Carrier Proteins
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GATA1 Transcription Factor
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GATA1 protein, human
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Glial Cell Line-Derived Neurotrophic Factor
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RNA, Small Interfering
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Repressor Proteins
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TP53 protein, human
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Tet O resistance protein, Bacteria
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Tumor Suppressor Protein p53
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tetracycline resistance-encoding transposon repressor protein
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Green Fluorescent Proteins
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DNA Polymerase II
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DNA Polymerase III
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Doxycycline