UNIT 4.32 Transfection of Cultured Primary Neurons via Nucleofection

  1. Manuel Zeitelhofer,
  2. John P. Vessey,
  3. Sabine Thomas,
  4. Michael Kiebler,
  5. Ralf Dahm

Published Online: 1 APR 2009

DOI: 10.1002/0471142301.ns0432s47

Current Protocols in Neuroscience

Current Protocols in Neuroscience

How to Cite

Zeitelhofer, M., Vessey, J. P., Thomas, S., Kiebler, M. and Dahm, R. 2009. Transfection of Cultured Primary Neurons via Nucleofection. Current Protocols in Neuroscience. 47:4.32:4.32.1–4.32.21.

Author Information

  1. Medical University of Vienna, Center for Brain Research, Vienna, Austria

Publication History

  1. Published Online: 1 APR 2009
  2. Published Print: APR 2009


Despite the development of various transfection methods, the transfection of post-mitotic cells, including neurons, poses a challenging task. Nucleofection, a specialized form of electroporation described in this unit, achieves high transfection efficiencies in primary mammalian neurons, such as hippocampal neurons, while simultaneously maintaining high cell viability. Therefore, it allows for biochemical analyses that rely on large numbers of transfected cells. The recently developed 96-well shuttle system described in this unit further permits the transfection of up to 96 different constructs in a single experiment. This opens up the possibility for large-scale experiments in primary neurons, such as shRNA-mediated knock-down of a wide range of target genes. Curr. Protoc. Neurosci. 47:4.32.1-4.32.21. © 2009 by John Wiley & Sons, Inc.


  • transfection;
  • nucleofection;
  • primary hippocampal neurons;
  • transfection efficiency;
  • expression plasmids;
  • RNAi knock-down