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Biolistic DNA Delivery pp 157–166Cite as

Biolistic Transfection of Neurons in Organotypic Brain Slices

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Part of the book series: Methods in Molecular Biology ((MIMB,volume 940))

Abstract

Transfection of postmitotic neurons is one of the most challenging goals in the field of gene delivery. Currently most procedures use dissociated cell cultures but organotypic slice preparations have significant advantages as an experimental system; they preserve the three-dimensional architecture and local environment of neurons, yet still allow access for experimental manipulations and observations. However exploring the effects of novel genes in these preparations requires a technique that can efficiently transfect cells deep into tissues. Here we show that biolistic transfection is an effective and straightforward technique with which to transfect such cells.

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References

  1. Felgner PL et al (1987) Lipofection: a highly efficient, lipid-mediated DNA-transfection procedure. Proc Natl Acad Sci USA 84:7413–7417

    Article  PubMed  CAS  Google Scholar 

  2. Monkkonen J, Urtti A (1998) Lipid fusion in oligonucleotide and gene delivery with cationic lipids. Adv Drug Deliv Rev 34:37–49

    Article  PubMed  CAS  Google Scholar 

  3. Ross PC, Hui SW (1999) Lipoplex size is a major determinant of in vitro lipofection efficiency. Gene Ther 6:651–659

    Article  PubMed  CAS  Google Scholar 

  4. Witkowski PT et al (2009) Gene gun-supported DNA immunisation of chicken for straightforward production of poxvirus-specific IgY antibodies. J Immunol Methods 341:146–153

    Article  PubMed  CAS  Google Scholar 

  5. O’Brien JA, Lummis SCR (2006) Biolistic transfection of neuronal cultures using a hand-held gene gun. Nat Protoc 1:977–981

    Article  PubMed  Google Scholar 

  6. O’Brien JA et al (2001) Modifications to the hand-held Gene Gun: improvements for in vitro biolistic transfection of organotypic neuronal tissue. J Neurosci Methods 112:57–64

    Article  PubMed  Google Scholar 

  7. Sato H et al (2000) In vivo gene gun-mediated DNA delivery into rodent brain tissue. Biochem Biophys Res Commun 270:163–170

    Article  PubMed  CAS  Google Scholar 

  8. Klein TM et al (1987) High-velocity microprojectiles for delivering nucleic acids into living cells. Nature 327:70–73

    Article  CAS  Google Scholar 

  9. Johnston SA (1990) Biolistic transformation: microbes to mice. Nature 346:776–770

    Article  PubMed  CAS  Google Scholar 

  10. Sanford JC, Smith FD, Russell JA (1993) Optimizing the biolistic process for different biological applications. Methods Enzymol 217:483–509

    Article  PubMed  CAS  Google Scholar 

  11. Cui Z, Mumper RJ (2003) The effect of co-administration of adjuvants with a nanoparticle-based genetic vaccine delivery system on the resulting immune responses. Eur J Pharm Biopharm 55:11–18

    Article  PubMed  CAS  Google Scholar 

  12. Cheng L, Ziegelhoffer PR, Yang NS (1993) In vivo promoter activity and transgene expression in mammalian somatic tissues evaluated by using particle bombardment. Proc Natl Acad Sci USA 90:4455–4459

    Article  PubMed  CAS  Google Scholar 

  13. O’Brien JA, Lummis SCR (2011) Nano-biolistics: a method of biolistic transfection of cells and tissues using a gene gun with novel nanometer-sized projectiles. BMC Biotechnol 11:66–76

    Article  PubMed  Google Scholar 

  14. Lo DC (2001) Neuronal transfection using particle-mediated gene transfer. Curr Protoc Neurosci. doi:10.1002/0471142301 UNIT3.15

    Google Scholar 

  15. Smith FD, Harpending PR, Sanford JC (1992) Biolistic transformation of prokaryotes: factors that affect biolistic transformation of very small cells. J Gen Microbiol 138:239–248

    Article  PubMed  CAS  Google Scholar 

  16. Barry MA, Singh RAK, Andersson HA (2003) Gene Gun technologies: applications for gene therapy and genetic immunization. In: Smyth-Templeton N (ed) Gene therapy: therapeutic mechanisms and strategies. Marcel Dekker Inc., New York, pp 263–285

    Chapter  Google Scholar 

  17. Donnelly JJ, Wahren B, Liu MA (2005) DNA vaccines: progress and challenges. J Immunol 175:633–639

    PubMed  CAS  Google Scholar 

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Acknowledgments

The work was supported by the Medical Research Council and the Wellcome Trust. S.C.R.L. holds a Wellcome Trust Senior Research Fellowship in Basic Biomedical Science.

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Authors and Affiliations

  1. Division of Neurobiology, MRC Laboratory of Molecular Biology, Cambridge, UK

    John A. O’Brien & Sarah C. R. Lummis

  2. Department of Biochemistry, University of Cambridge, Cambridge, UK

    Sarah C. R. Lummis

Authors
  1. John A. O’Brien
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  2. Sarah C. R. Lummis
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Corresponding author

Correspondence to Sarah C. R. Lummis .

Editor information

Editors and Affiliations

  1. Ganzimmun Diagnostics AG, Hans-Böckler-Str. 109, Mainz, 55128, Germany

    Stephan Sudowe

  2. University Medical Center, Department of Dermatology, Johannes Gutenberg-University, Obere Zahlbacher Str. 63, Mainz, 55131, Germany

    Angelika B. Reske-Kunz

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© 2013 Springer Science+Business Media, LLC

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O’Brien, J.A., Lummis, S.C.R. (2013). Biolistic Transfection of Neurons in Organotypic Brain Slices. In: Sudowe, S., Reske-Kunz, A. (eds) Biolistic DNA Delivery. Methods in Molecular Biology, vol 940. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-110-3_13

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  • DOI: https://doi.org/10.1007/978-1-62703-110-3_13

  • Published:

  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-62703-109-7

  • Online ISBN: 978-1-62703-110-3

  • eBook Packages: Springer Protocols

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