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Biolistic transfection of neuronal cultures using a hand-held gene gun

Nature Protocols volume 1, pages 977–981 (2006)Cite this article

Abstract

Biolistic transfection is a technique in which subcellular-sized particles coated with DNA are accelerated to high velocity to propel them into cells. This method is applicable to tissues, cells and organelles, and can be used for both in vitro and in vivo transformations; with the right equipment, it is simple, rapid and efficient. Here we provide a detailed protocol for biolistic transfection of plasmids into cultured human embryonic kidney (HEK) 293 cells and organotypic brain slices using a hand-held gene gun. There are three major steps: (i) coating microcarriers with DNA, (ii) transferring the microcarriers into a cartridge to make a 'bullet', and (iii) firing the DNA-coated microcarriers into cells using a pulse of helium gas. The method can be readily adapted to other cell types and tissues. The protocol can be completed in 1–2 h.

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Figure 1: Helios gene gun.
Figure 2: Using the gene gun.
Figure 3: HEK293 cells expressing EYFP.
Figure 4: A neuron in a cerebellar brain slice labeled with EYFP.
Figure 5: A neuron in a brain slice expressing two protein subunits.

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Acknowledgements

This 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. Neurobiology Division, Medical Research Council Laboratory of Molecular Biology, Hills Road, Cambridge, CB2 2QH, UK

    John A O'Brien & Sarah C R Lummis

  2. Department of Biochemistry, University of Cambridge, Cambridge, CB2 1QW, UK

    Sarah C R Lummis

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  1. John A O'Brien
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  2. Sarah C R Lummis
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Correspondence to Sarah C R Lummis.

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O'Brien, J., Lummis, S. Biolistic transfection of neuronal cultures using a hand-held gene gun. Nat Protoc 1, 977–981 (2006). https://doi.org/10.1038/nprot.2006.145

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