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Biolistic Transfection of Human Embryonic Kidney (HEK) 293 Cells

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Biolistic DNA Delivery

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

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Abstract

Due to its excellent transfectability, the human embryonic kidney (HEK) 293 cell line is widely used as an in vitro model system for transfection experiments. Particle bombardment, or biolistics technology, provides a physical transfection approach that can deliver transgene materials efficiently into many different cell lines. Transfection of 293 cells by gene gun, allows examination of transgene expression in epithelial cells, as well as studies concerning a variety of questions in neurobiology. The present study of transfection of HEK 293 cells by biolistics technology uses the plasmids gWIZ-luc encoding luciferase and gWIZ-GFP encoding green fluorescence protein (GFP) as model transgenes. This system can be routinely used at varying bombarding conditions that can be adjusted according to experimental requirements and purpose, such as gene gun helium pressure, the sizes and the amount of the gold particles and the length of the spacer. The results obtained show that the Bio-Rad spacer for the gene gun should be optimized for travel distance and spreading of gold particles over a relatively small area, when used for biolistic transfection of cells dispersed in multi-well plate.

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References

  1. HEK cell-Wikipedia, http://en.wikipedia.org/wiki/HEK_cell

  2. Graham FL et al (1977) Characteristics of a human cell line transformed by DNA from human adenovirus type 5. J Gen Virol 36:59–74

    Article  PubMed  CAS  Google Scholar 

  3. Louis N, Evelegh C, Graham FL (1997) Cloning and sequencing of the cellular viral junctions from the human adenovirus type 5 transformed 293 cell line. Virology 233:423–429

    Article  PubMed  CAS  Google Scholar 

  4. Neumann E et al (1982) Gene transfer into mouse lyoma cells by electroporation in high electric fields. EMBO J 1:841–845

    PubMed  CAS  Google Scholar 

  5. Li XW et al (2003) Sustained expression in mammalian cells with DNA complexed with chitosan nanoparticles. Biochim Biophys Acta 1630:7–18

    Article  PubMed  CAS  Google Scholar 

  6. Uchida M et al (2009) Transfection by particle bombardment: delivery of plasmid DNA into mammalian cells using gene gun. Biochim Biophys Acta 1790:754–764

    Article  PubMed  CAS  Google Scholar 

  7. Kim H, Park Y (2002) Effect of lipid compositions on gene transfer into 293 cells using Sendai F/HN-virosomes. J Biochem Mol Biol 35:459–464

    Article  PubMed  CAS  Google Scholar 

  8. Corsi K et al (2003) Mesenchymal stem cells, MG63 and HEK293 transfection using chitosan-DNA nanoparticles. Biomaterials 24:1255–1264

    Article  PubMed  CAS  Google Scholar 

  9. Shaw G et al (2002) Preferential transformation of human neuronal cells by human adenoviruses and the origin of HEK 293 cells. FASEB J 16:869–71

    PubMed  CAS  Google Scholar 

  10. 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 

  11. Thomas P, Smart TG (2005) HEK293 cell line: a vehicle for the expression of recombinant proteins. J Pharmacol Toxicol Methods 51:187–200

    Article  PubMed  CAS  Google Scholar 

  12. Johnston SA, Tang DC (1994) Gene gun transfection of animal cells and genetic immunization. Methods Cell Biol 43:353–365

    Article  PubMed  CAS  Google Scholar 

  13. Yang NS et al (1990) In vivo and in vitro gene transfer to mammalian somatic cells by particle bombardment. Proc Natl Acad Sci USA 87:9568–9572

    Article  PubMed  CAS  Google Scholar 

  14. Williams RS et al (1991) Introduction of foreign genes into tissues of living mice by DNA-coated microprojectiles. Proc Natl Acad Sci USA 88:2726–2730

    Article  PubMed  CAS  Google Scholar 

  15. 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 

  16. 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 

  17. 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 

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Acknowledgments

This work was supported by the BBSRC grant BB/D014204/1.

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Correspondence to H. Oya Alpar .

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Li, X., Uchida, M., Alpar, H.O., Mertens, P. (2013). Biolistic Transfection of Human Embryonic Kidney (HEK) 293 Cells. 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_10

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  • DOI: https://doi.org/10.1007/978-1-62703-110-3_10

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  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-62703-109-7

  • Online ISBN: 978-1-62703-110-3

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