Gene transfer engineering for astrocyte-specific silencing in the CNS

• Published in: Gene therapy Vol. 22; no. 10; pp. 830 - 839
• Main Authors: Merienne, N, Delzor, A, Viret, A, Dufour, N, Rey, M, Hantraye, P, Déglon, N
• Format: Journal Article
• Language: English
• Published: England Nature Publishing Group 01.10.2015
• Subjects: Animals; Astrocytes; Astrocytes - physiology; Central nervous system; Central Nervous System - cytology; Central Nervous System - physiology; Central nervous system diseases; Gene Expression Regulation; Gene Silencing; Gene therapy; Gene transfer; Gene Transfer Techniques; Genes; Genetic engineering; Genetic transformation; Genetic Vectors; Glutamate-ammonia ligase; Glutamine; Health aspects; Lentivirus; Ligases; Mice; Mice, Transgenic; MicroRNA; MicroRNAs; miRNA; Nervous system; Neurons; RNA; RNA processing; RNA, Small Interfering; Tetracycline; Tetracyclines; Therapeutic applications; France; Switzerland; Germany
• ISSN: 0969-7128; 1476-5462
• DOI: 10.1038/gt.2015.54

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.