Skip to main content
SpringerLink
Log in

Transfection of Neuronal Cultures

  • Protocol
  • First Online:
Neuronal Cell Culture

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2311))

  • 2182 Accesses

Abstract

Efficient transfection of genes into the neurons is a crucial step for the study of neuronal cell biology and functions. These include but not limited to investigating gene function by overexpression of target proteins via expression plasmids and knocking down the expression levels of neuronal genes by RNA interference (RNAi). In addition, reporter gene constructs are widely used to investigate the promoter activities of neuronal genes. Numerous transfection techniques have been established to deliver genes into the cells. However, efficient transfection of postmitotic cells, including neurons, still remains a challenging task. Here, we overview the advantages and disadvantages of various techniques for the transfection of primary neurons, and provide an optimized protocol for FuGENE-6 (Promega) which allows for a suitable transfection efficiency of primary neuronal cultures.

This is a preview of subscription content, log in via an institution to check access.

Access this chapter

Log in via an institution
Protocol
USD 49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 89.00
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 119.99
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD 199.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

References

  1. http://www.promega.com/resources/product-guides-and-selectors/protocols-and-applications-guide/transfection/

  2. Zeitelhofer M, John P, Vessey JP, Thomas S, Kiebler M, Dahm R (2009) Transfection of cultured primary neurons via nucleofection. Curr Protoc Neurosci Chapter 4:Unit4.32

    PubMed  Google Scholar 

  3. Karra D, Dahm R (2010) Transfection Techniques for Neuronal Cells. J Neurosci 30(18):6171–6177

    Article  CAS  Google Scholar 

  4. Washbourne P, McAllister AK (2002) Techniques for gene transfer into neurons. Curr Opin Neurobiol 12:566–573

    Article  CAS  Google Scholar 

  5. Dib-Hajj SD, Choi JS, Macala LJ, Tyrrell L, Black JA, Cummins TR, Waxman SG (2009) Transfection of rat or mouse neurons by biolistics or electroporation. Nat Protoc 4:1118–1126

    Article  CAS  Google Scholar 

  6. Gresch O, Engel FB, Nesic D, Tran TT, England HM, Hickman ES, Korner I, Gan L, Chen S, Castro-Obregon S, Hammermann R, Wolf J, Müller-Hartmann H, Nix M, Siebenkotten G, Kraus G, Lun K (2004) New non-viral method for gene transfer into primary cells. Methods 33:151–163

    Article  CAS  Google Scholar 

  7. Gartner A, Collin L, Lalli G (2006) Nucleofection of primary neurons. Methods Enzymol 406:374–388

    Article  CAS  Google Scholar 

  8. Chen C, Okayama H (1987) High-efficiency transformation of mammalian cells by plasmid DNA. Mol Cell Biol 7:2745–2752

    Article  CAS  Google Scholar 

  9. Yu LY, Arumäe U (2008) Survival assay of transiently transfected dopaminergic neurons. J Neurosci Methods 169(1):8–15

    Article  CAS  Google Scholar 

  10. Gabellini N, Minozzi MC, Leon A, Dal Toso R (1992) Nerve growth factor transcriptional control of c-fos promoter transfected in cultured spinal sensory neurons. J Cell Biol 118(1):131–138

    Article  CAS  Google Scholar 

  11. Goetze B, Grunewald B, Baldassa S, Kiebler M (2004) Chemically controlled formation of a DNA/calcium phosphate coprecipitate: application for transfection of mature hippocampal neurons. J Neurobiol 60:517–525

    Article  CAS  Google Scholar 

  12. Felgner PL, Gadek TR, Holm M, Roman R, Chan HW, Wenz M, Northrop JP, Ringold GM, Danielsen M (1987) Lipofection: a highly efficient, lipid-mediated DNA-transfection procedure. Proc Natl Acad Sci 84:7413–7417

    Article  CAS  Google Scholar 

  13. Felgner JH, Kumar R, Sridhar CN, Wheeler CJ, Tsai YJ, Border R, Ramsey P, Martin M, Felgner PL (1994) Enhanced gene delivery and mechanism studies with a novel series of cationic lipid formulations. J Biol Chem 269(4):2550–2561

    Article  CAS  Google Scholar 

  14. Wiesenhofer B, Humpel C (2000) Lipid-mediated gene transfer into primary neurons using FuGene: comparison to C6 Glioma cells and primary glia. Exp Neurol 164:38–44

    Article  CAS  Google Scholar 

  15. Ohki EC, Tilkins ML, Ciccarone VC, Price PJ (2001) Improving the transfection efficiency of post-mitotic neurons. J Neurosci Methods 112:95–99

    Article  CAS  Google Scholar 

Download references

Acknowledgments

This work was supported by NIH grants awarded to IKS.

Author information

Authors and Affiliations

  1. Department of Neuroscience, Lewis Katz School of Medicine, Temple University, Philadelphia, PA, USA

    Ilker Kudret Sariyer

Authors
  1. Ilker Kudret Sariyer
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Ilker Kudret Sariyer .

Editor information

Editors and Affiliations

  1. College of Science and Technology, Temple University, Philadelphia, PA, USA

    Shohreh Amini

  2. Department of Neuroscience, Lewis Katz School of Medicine, Temple University, Philadelphia, PA, USA

    Martyn K. White

Rights and permissions

Reprints and permissions

Copyright information

© 2021 Springer Science+Business Media, LLC, part of Springer Nature

About this protocol

Check for updates. Verify currency and authenticity via CrossMark

Cite this protocol

Sariyer, I.K. (2021). Transfection of Neuronal Cultures. In: Amini, S., White, M.K. (eds) Neuronal Cell Culture. Methods in Molecular Biology, vol 2311. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-1437-2_10

Download citation

  • DOI: https://doi.org/10.1007/978-1-0716-1437-2_10

  • Published:

  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-1436-5

  • Online ISBN: 978-1-0716-1437-2

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics

Search

Navigation