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.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
HEK cell-Wikipedia, http://en.wikipedia.org/wiki/HEK_cell
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
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
Neumann E et al (1982) Gene transfer into mouse lyoma cells by electroporation in high electric fields. EMBO J 1:841–845
Li XW et al (2003) Sustained expression in mammalian cells with DNA complexed with chitosan nanoparticles. Biochim Biophys Acta 1630:7–18
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
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
Corsi K et al (2003) Mesenchymal stem cells, MG63 and HEK293 transfection using chitosan-DNA nanoparticles. Biomaterials 24:1255–1264
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
O’Brien JA, Lummis SCR (2006) Biolistic transfection of neuronal cultures using a hand-held gene gun. Nat Protoc 1:977–981
Thomas P, Smart TG (2005) HEK293 cell line: a vehicle for the expression of recombinant proteins. J Pharmacol Toxicol Methods 51:187–200
Johnston SA, Tang DC (1994) Gene gun transfection of animal cells and genetic immunization. Methods Cell Biol 43:353–365
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
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
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
Sato H et al (2000) In vivo gene gun-mediated DNA delivery into rodent brain tissue. Biochem Biophys Res Commun 270:163–170
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
Acknowledgments
This work was supported by the BBSRC grant BB/D014204/1.
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2013 Springer Science+Business Media, LLC
About this protocol
Cite this protocol
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
Download citation
DOI: https://doi.org/10.1007/978-1-62703-110-3_10
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