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Biolistic Transfection of Human Embryonic Kidney (HEK) 293 Cells

  • Xiongwei Li
  • Masaki Uchida
  • H. Oya AlparEmail author
  • Peter Mertens
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 940)

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.

Key words

HEK 293 cells Gene Transfection Biolistics Gene gun Gold particles Confocal microscope Plasmid DNA 

Notes

Acknowledgments

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

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Copyright information

© Springer Science+Business Media, LLC 2013

Authors and Affiliations

  • Xiongwei Li
    • 1
  • Masaki Uchida
    • 1
    • 2
  • H. Oya Alpar
    • 3
    Email author
  • Peter Mertens
    • 4
  1. 1.Department of Pharmaceutics, School of PharmacyUniversity of LondonLondonUK
  2. 2.Josai UniversitySakadoJapan
  3. 3.Division of Clinical Microbiology, Department of Pharmaceutics, School of PharmacyUniversity of LondonLondonUK
  4. 4.Institute for Animal Health, Pirbright LaboratoryPirbrightUK

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