Summary
The ability to manipulate RNAi in cultured mammalian cells has provided scientists with a very powerful tool to influence gene expression. Neurons represent a cell type that initially displayed resistance to transduction by siRNAs or shRNA, when attempting to silence expression of endogenous genes. However, the development of lentiviral systems with that goal has facilitated the exogenous manipulation of RNAi in these postmitotic cells. Lentiviral-mediated RNAi experiments in cultured mammalian neurons can be designed to address a wide variety of biological questions or to test potential therapeutic hairpins before moving to treatment trials in vivo. We provide a practical approach to accomplish siRNA-mediated silencing of the disease-linked protein torsinA in primary neuronal cultures through the generation of lentiviral vectors expressing shRNAs.
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Acknowledgments
Research on RNAi was funded by the Dystonia Medical Research Foundation (P. G.-A.) and NIH/NINDS (S. Q. H. and P. G.-A.). We are grateful to all the members of the Davidson, Gonzalez, and Paulson laboratories and the Gene Transfer Vectors Core at The University of Iowa for all the hard work carried out to develop and optimize the protocols described here.
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© 2008 Humana Press, a part of Springer Science+Business Media, LLC
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Harper, S.Q., Gonzalez-Alegre, P. (2008). Lentivirus-Mediated RNA Interference in Mammalian Neurons. In: Barik, S. (eds) RNAi. Methods in Molecular Biology™, vol 442. Humana Press. https://doi.org/10.1007/978-1-59745-191-8_8
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DOI: https://doi.org/10.1007/978-1-59745-191-8_8
Publisher Name: Humana Press
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