96-Well electroporation method for transfection of mammalian central neurons

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

Research output: Contribution to journalArticle

25 Citations (Scopus)

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
Pages (from-to)619-624
Number of pages6
JournalBioTechniques
Volume41
Issue number5
DOIs
StatePublished - Nov 1 2006

Fingerprint

Electroporation
Neurons
Transfection
Plasmids
Poisons
Neurology
Virus Diseases
Gene expression
Vertebrates
Screening
Central Nervous System
Phenotype
Lipids
Gene Expression
Costs and Cost Analysis
Costs

ASJC Scopus subject areas

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

Cite this

96-Well electroporation method for transfection of mammalian central neurons. / Buchser, William J.; Pardinas, Jose R.; Shi, Yan; Bixby, John; Lemmon, Vance.

In: BioTechniques, Vol. 41, No. 5, 01.11.2006, p. 619-624.

Research output: Contribution to journalArticle

Buchser, William J. ; Pardinas, Jose R. ; Shi, Yan ; Bixby, John ; Lemmon, Vance. / 96-Well electroporation method for transfection of mammalian central neurons. In: BioTechniques. 2006 ; Vol. 41, No. 5. pp. 619-624.
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