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Local gene knockdown in the brain using viral-mediated RNA interference

Nature Medicine volume 9, pages 1539–1544 (2003)Cite this article

Abstract

Conditional mutant techniques that allow spatial and temporal control over gene expression can be used to create mice with restricted genetic modifications. These mice serve as powerful disease models in which gene function in adult tissues can be specifically dissected. Current strategies for conditional genetic manipulation are inefficient, however, and often lack sufficient spatial control. Here we use viral-mediated RNA interference (RNAi) to generate a specific knockdown of Th, the gene encoding the dopamine synthesis enzyme tyrosine hydroxylase, within midbrain neurons of adult mice. This localized gene knockdown resulted in behavioral changes, including a motor performance deficit and reduced response to a psychostimulant. These results underscore the potential of using viral-mediated RNAi for the rapid production and testing of new genetic disease models. Similar strategies may be used in other model species, and may ultimately find applications in human gene therapy.

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Figure 1: Viral-mediated RNAi reduces tyrosine hydroxylase in the substantia nigra compacta.
Figure 2: Time-course analysis of viral-mediated Th knockdown.
Figure 3: AAV injection in the midbrain results in efficient infection of defined brain regions.
Figure 4: Infection of VTA neurons reduces tyrosine hydroxylase in the nucleus accumbens.
Figure 5: Regional Th knockdown results in behavioral deficits.

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Acknowledgements

We thank D. Turner for providing the U6 promoter-containing vector, L. Perrotti for rotarod assay protocols, E. Kim and L. Leverich for help with data collection, and C. Bolaños for advice on data analysis.

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  1. Department of Psychiatry, The University of Texas Southwestern Medical Center at Dallas, 5323 Harry Hines Blvd., Dallas, 75390–9070, Texas, USA

    Jonathan D Hommel, Robert M Sears, Dan Georgescu, Diana L Simmons & Ralph J DiLeone

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  1. Jonathan D Hommel
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  2. Robert M Sears
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Correspondence to Ralph J DiLeone.

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Hommel, J., Sears, R., Georgescu, D. et al. Local gene knockdown in the brain using viral-mediated RNA interference. Nat Med 9, 1539–1544 (2003). https://doi.org/10.1038/nm964

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