Artículo
Dual-promoter lentiviral vectors for constitutive and regulated gene expression in neurons
Autor/es | Gascón, Sergio
Páez Gómez, Juan A. Díaz-Guerra, Margarita Scheiffele, Peter Gómez Scholl, Francisco Manuel |
Departamento | Universidad de Sevilla. Departamento de Fisiología Médica y Biofísica |
Fecha de publicación | 2008-02-15 |
Fecha de depósito | 2024-07-31 |
Publicado en |
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Resumen | Gene transfer methods for efficient co-expression of exogenous proteins in neurons are crucial tools towards the understanding of the molecular basis of the central nervous system. Lentiviruses are retroviral vectors that ... Gene transfer methods for efficient co-expression of exogenous proteins in neurons are crucial tools towards the understanding of the molecular basis of the central nervous system. Lentiviruses are retroviral vectors that can transduce a wide variety of cells including differentiated neurons. In this work, we have generated lentiviral vectors containing dual promoters that allow efficient co-expression of exogenous proteins in neurons. We show that insertion of two copies of a human synapsin promoter/WPRE cassette in a single lentiviral vector directs robust co-expression of cDNAs in cultured neurons, while excluding expression in the surrounding glial cells. Furthermore, insertion of the tetracycline-inducible system (Tet-off) controlled by the synapsin promoter results in tightly regulated expression of EGFP when used as a transgene in cultured neurons. Transduction of primary neurons with this inducible system leads to a 100-fold increase in EGFP mRNA levels in the absence of doxycycline. In transduced cultures, EGFP transcription is inhibited within 24 h upon addition of doxycycline. The viral systems we developed here provide neuron-specific and regulated expression mediated by single lentiviral vectors and will prove valuable tools for the study of neuronal function. |
Cita | Gascón, S., Páez Gómez, J.A., Díaz-Guerra, M., Scheiffele, P. y Gómez Scholl, F.M. (2008). Dual-promoter lentiviral vectors for constitutive and regulated gene expression in neurons. Journal of Neuroscience Methods, 168 (1), 104-112. https://doi.org/10.1016/j.jneumeth.2007.09.023. |
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Dual promoter lentiviral vectors ... | 202.1Kb | [PDF] | Ver/ | Versión aceptada / Accepted version |