96-Well electroporation method for transfection of mammalian central neurons

William J. Buchser, Jose R. Pardinas, Yan Shi, John L. Bixby, Vance P. Lemmon

Research output: Contribution to journalArticle

25 Scopus citations

Abstract

Manipulating gene expression in primary neurons has been a goal for many scientists for over 20 years. Vertebrate central nervous system neurons are classically difficult to transfect. Most lipid reagents are inefficient and toxic to the cells, and time-consuming methods such as viral infections are often required to obtain better efficiencies. We have developed an efficient method for the transfection of cerebellar granule neurons and hippocampal neurons with standard plasmid vectors. Using 96-well electroporation plates, square-wave pulses can introduce 96 different plasmids into neurons in a single step. The procedure results in greater than 20% transfection efficiencies and requires only simple solutions of nominal cost. In addition to enabling the rapid optimization of experimental protocols with multiple parameters, this procedure enables the use of high content screening methods to characterize neuronal phenotypes.

Original languageEnglish (US)
Pages (from-to)619-624
Number of pages6
JournalBioTechniques
Volume41
Issue number5
DOIs
StatePublished - Nov 2006

ASJC Scopus subject areas

  • Biotechnology
  • Biochemistry, Genetics and Molecular Biology(all)

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