title: Gene transfer engineering for astrocyte-specific silencing in the CNS.
abstract: Cell-type-specific gene silencing is critical to understand cell functions in normal and pathological conditions, in particular in the brain where strong cellular heterogeneity exists. Molecular engineering of lentiviral vectors has been widely used to express genes of interest specifically in neurons or astrocytes. However, we show that these strategies are not suitable for astrocyte-specific gene silencing due to the processing of small hairpin RNA (shRNA) in a cell. Here we develop an indirect method based on a tetracycline-regulated system to fully restrict shRNA expression to astrocytes. The combination of Mokola-G envelope pseudotyping, glutamine synthetase promoter and two distinct microRNA target sequences provides a powerful tool for efficient and cell-type-specific gene silencing in the central nervous system. We anticipate our vector will be a potent and versatile system to improve the targeting of cell populations for fundamental as well as therapeutic applications.
authors.affiliation: Laboratory of Cellular and Molecular Neurotherapies (LCMN), Department of Clinical Neurosciences (DNC), Lausanne University Hospital (CHUV), Lausanne, Switzerland.|LCMN, Neuroscience Research Center (CRN), Lausanne University Hospital, Lausanne, Switzerland.
authors.forename: N
authors.initials: N
authors.lastname: Merienne
authors.affiliation: Commissariat à l'Energie Atomique et aux Energies Alternatives (CEA), Institute of Biomedical Imaging (I2BM), Molecular Imaging Research Center (MIRCen), Fontenay-aux-Roses, France.|CNRS-CEA URA2210, Fontenay-aux-Roses, France.
authors.forename: A
authors.initials: A
authors.lastname: Delzor
authors.affiliation: Laboratory of Cellular and Molecular Neurotherapies (LCMN), Department of Clinical Neurosciences (DNC), Lausanne University Hospital (CHUV), Lausanne, Switzerland.|LCMN, Neuroscience Research Center (CRN), Lausanne University Hospital, Lausanne, Switzerland.
authors.forename: A
authors.initials: A
authors.lastname: Viret
authors.affiliation: Commissariat à l'Energie Atomique et aux Energies Alternatives (CEA), Institute of Biomedical Imaging (I2BM), Molecular Imaging Research Center (MIRCen), Fontenay-aux-Roses, France.|CNRS-CEA URA2210, Fontenay-aux-Roses, France.
authors.forename: N
authors.initials: N
authors.lastname: Dufour
authors.affiliation: Laboratory of Cellular and Molecular Neurotherapies (LCMN), Department of Clinical Neurosciences (DNC), Lausanne University Hospital (CHUV), Lausanne, Switzerland.|LCMN, Neuroscience Research Center (CRN), Lausanne University Hospital, Lausanne, Switzerland.
authors.forename: M
authors.initials: M
authors.lastname: Rey
authors.affiliation: Commissariat à l'Energie Atomique et aux Energies Alternatives (CEA), Institute of Biomedical Imaging (I2BM), Molecular Imaging Research Center (MIRCen), Fontenay-aux-Roses, France.|CNRS-CEA URA2210, Fontenay-aux-Roses, France.
authors.forename: P
authors.initials: P
authors.lastname: Hantraye
authors.affiliation: Laboratory of Cellular and Molecular Neurotherapies (LCMN), Department of Clinical Neurosciences (DNC), Lausanne University Hospital (CHUV), Lausanne, Switzerland.|LCMN, Neuroscience Research Center (CRN), Lausanne University Hospital, Lausanne, Switzerland.
authors.forename: N
authors.initials: N
authors.lastname: Déglon
mesh_terms: D000818:Animals; D001253:Astrocytes / Q000502:physiology*; D002490:Central Nervous System / Q000166:cytology / Q000502:physiology; D005786:Gene Expression Regulation; D020868:Gene Silencing; D018014:Gene Transfer Techniques; D005822:Genetic Vectors; D016086:Lentivirus; D051379:Mice; D008822:Mice, Transgenic; D034741:RNA, Small Interfering; D013752:Tetracycline
chemical_list: D034741:RNA, Small Interfering; D013752:Tetracycline