Optimal promoter usage for lentiviral vector-mediated transduction of cultured central nervous system cells

Mingjie Li, Nada Husic, Ying Lin, Heather Christensen, Ibrahim Malik, Sally McIver, Christine M LaPash Daniels, David A. Harris, Paul T. Kotzbauer, Mark P. Goldberg, B. Joy Snider

Research output: Contribution to journalArticle

41 Scopus citations

Abstract

Lentiviral vectors transduce both dividing and non-dividing cells and can support sustained expression of transgenes. These properties make them attractive for the transduction of neurons and other neural cell types in vitro and in vivo. Lentiviral vectors can be targeted to specific cell types by using different promoters in the lentiviral shuttle vector. Even with identical constructs, however, levels of expression can vary significantly in different types of neurons and different culture preparations; expression levels in the same neuronal subtypes can be very different in primary cell culture and in vivo. We systematically assessed the ability of different promoters to direct expression of foreign transgenes in primary murine neocortical neurons, cerebellar granule cells and in undifferentiated and differentiated neuroblastoma cells. In primary cortical neurons, constructs using the ubiquitin C promoter directed the highest level of transgene expression; the phosphoglycerate kinase (PGK) promoter also directed robust transgene expression, while the cytomegalovirus (CMV) and MND (a synthetic promoter that contains the U3 region of a modified MoMuLV LTR with myeloproliferative sarcoma virus enhancer) promoters resulted in the expression of the transgenes in only limited number of neurons. In contrast, in cerebellar granule cells and in differentiated SH-SY5Y neuroblastoma cultures, the CMV promoter directed the most robust transgene expression. There was similar variability in transgene expression directed by these promoters in primary cultures of oligodendrocytes and astrocytes. These findings may prove useful in the design of lentiviral vectors for use in cell culture models of the nervous system.

Original languageEnglish (US)
Pages (from-to)56-64
Number of pages9
JournalJournal of Neuroscience Methods
Volume189
Issue number1
DOIs
StatePublished - May 1 2010

Keywords

  • Astrocyte
  • Lentiviral vector
  • Neuron
  • Promoter

ASJC Scopus subject areas

  • Neuroscience(all)

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    Li, M., Husic, N., Lin, Y., Christensen, H., Malik, I., McIver, S., Daniels, C. M. L., Harris, D. A., Kotzbauer, P. T., Goldberg, M. P., & Snider, B. J. (2010). Optimal promoter usage for lentiviral vector-mediated transduction of cultured central nervous system cells. Journal of Neuroscience Methods, 189(1), 56-64. https://doi.org/10.1016/j.jneumeth.2010.03.019