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Manipulation of Gene Expression in the Central Nervous System with Lentiviral Vectors

  • Binggui Sun
  • Li GanEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 670)

Abstract

Viral vector-mediated gene transfer is widely used to manipulate gene expression (overexpression or knock down) in cultures and in different tissues of animals. Vectors based on lentiviruses have particularly useful features. Lentiviral vectors mediate gene transfer into any neuronal cell types and induce sustained expression without significant immune responses after delivery into the nervous system. Lentivirus-mediated expression of therapeutic genes has led to long-term treatment of animal models of neurological disorders, such as spinal injury, Parkinson’s disease, Huntington’s disease, and Alzheimer’s disease. Here, we describe the preparation and purification of lentiviral vectors and methods of lentiviral infection in primary neural cultures and in brain regions of interest by stereotaxic injection.

Key words

Lentiviral vector Gene expression Primary neural culture Stereotaxic injection Central nervous system 

Notes

Acknowledgments

The authors thank Yungui Zhou for discussion and advice on this chapter. This work was supported in part by NIH grant (AG024447) and a pilot project grant from the UCSF Alzheimer’s Disease Research Center (to L.G.).

References

  1. 1.
    Zufferey, R., Dull, T., Mandel, R. J., Bukovsky, A., Quiroz, D., Naldini, L., et al. (1998) Self-inactivating lentivirus vector for safe and efficient in vivo gene delivery. J. Virol. 72, 9873–9880.PubMedGoogle Scholar
  2. 2.
    Baekelandt, V., Claeys, A., Eggermont, K., Lauwers, E., De Strooper, B., Nuttin, B., et al. (2002) Characterization of lentiviral vector-mediated gene transfer in adult mouse brain. Hum. Gene Ther. 13, 841–853.PubMedCrossRefGoogle Scholar
  3. 3.
    Palfi, S., Leventhal, L., Chu, Y., Ma, S. Y., Emborg, M., Bakay, R., et al. (2002) Lentivirally delivered glial cell line-derived neurotrophic factor increases the number of striatal dopaminergic neurons in primate models of nigrostriatal degeneration. J. Neurosci. 22, 4942–4954.PubMedGoogle Scholar
  4. 4.
    Lois, C., Hong, E. J., Pease, S., Brown, E. J., Baltimore, D. (2002) Germline transmission and tissue-specific expression of transgenes delivered by lentiviral vectors. Science 295, 868–872.PubMedCrossRefGoogle Scholar
  5. 5.
    Paxinos, G. and Franklin, K. B. J. (ed.) (2001) The mouse brain in stereotaxic coordinates. Academic Press, San Diego, CA.Google Scholar
  6. 6.
    Chen, J., Zhou, Y., Mueller-Steiner, S., Chen, L. F., Kwon, H., Yi, S., et al. (2005) SIRT1 Protects against microglia-dependent amyloid-beta toxicity through inhibiting NF-kappaB signaling. J. Biol. Chem. 280, 40364–40374.PubMedCrossRefGoogle Scholar
  7. 7.
    Mueller-Steiner, S., Zhou, Y., Arai, H., Roberson, E. D., Sun, B., Chen, J., et al. (2006) Antiamyloidogenic and neuroprotective functions of cathepsin B: Implications for Alzheimer’s disease. Neuron 51, 703–714.PubMedCrossRefGoogle Scholar

Copyright information

© Humana Press 2010

Authors and Affiliations

  1. 1.Department of Neurology, Gladstone Institute of Neurological DiseaseUniversity of California, San FranciscoSan FranciscoUSA

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