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Highly efficient molecular delivery into mammalian cells using carbon nanotube spearing

Nature Methods volume 2, pages 449–454 (2005)Cite this article

Abstract

Introduction of exogenous DNA into mammalian cells represents a powerful approach for manipulating signal transduction. The available techniques, however, are limited by low transduction efficiency and low cell viability after transduction. Here we report a highly efficient molecular delivery technique, named nanotube spearing, based on the penetration of nickel-embedded nanotubes into cell membranes by magnetic field driving. DNA plasmids containing the enhanced green fluorescent protein (EGFP) sequence were immobilized onto the nanotubes, and subsequently speared into targeted cells. We have achieved an unprecedented high transduction efficiency in Bal17 B-lymphoma, ex vivo B cells and primary neurons with high viability after transduction. This technique may provide a powerful tool for highly efficient gene transfer into a variety of cells, especially the hard-to-transfect cells.

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Figure 1: A two-step procedure of nanotube spearing.
Figure 2: Nanotube spearing in MCF-7 cells.
Figure 3: Transduction of pEGFP-c1 in Bal17 cells by nanotube spearing.
Figure 4: EGFP transduction in primary cultured cells by nanotube spearing.

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Acknowledgements

The work performed at Boston College is partly supported by the US Department of Energy (DE-FG02-00ER45805), National Science Foundation (NIRT 0304506), and National Institutes of Health (Public Health Service Grant AI-34586). We thank D. Burgess (Boston College, Department of Biology) for the EGFP expressing plasmid. We also thank members of the Burgess' laboratory and Department of Biology for allowing us to use the Confocal Microscopy Facility.

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Authors and Affiliations

  1. NanoLab, Inc., Newton, 02458, Massachusetts, USA

    Dong Cai, Zhongping Huang & David Carnahan

  2. Department of Biology, Boston College, Chestnut Hill, 02467, Massachusetts, USA

    Jennifer M Mataraza & Thomas C Chiles

  3. Department of Neurology, Laboratory of Cellular Neurology, Massachusetts General Hospital and Harvard Medical School, Charlestown, 02129, Massachusetts, USA

    Zheng-Hong Qin

  4. Department of Pharmacology, Soochow University School of Medicine, 215007, Suzhou, China

    Zheng-Hong Qin

  5. Department of Physics, Boston College, Chestnut Hill, 02467, Massachusetts, USA

    Jianyu Huang, Kris Kempa & Zhifeng Ren

Authors
  1. Dong Cai
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  2. Jennifer M Mataraza
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  4. Zhongping Huang
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  5. Jianyu Huang
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  6. Thomas C Chiles
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  7. David Carnahan
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  8. Kris Kempa
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  9. Zhifeng Ren
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Corresponding author

Correspondence to Dong Cai.

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Competing interests

Nanolab, Inc. is planning to commercialize the technique described in this article.

Supplementary information

Supplementary Fig. 1

Carbon nanotube and Ni particle morphology. (PDF 210 kb)

Supplementary Fig. 2

Cell mortality after nanotube spearing. (PDF 60 kb)

Supplementary Fig. 3

The layers of graphene sheets and/or amorphous carbon that enclose the Ni particle in the carbon nanotubes. (PDF 82 kb)

Supplementary Fig. 4

The Ni particle deprived nanotube. (PDF 93 kb)

Supplementary Fig. 5

Nuclei staining with DAPI in cortical neurons 72 hours after spearing. (PDF 177 kb)

Supplementary Fig. 6

EGFP transfection with Lipofectamine 2000 in Bal17 cells. (PDF 279 kb)

Supplementary Fig. 7

EGFP transfection with Lipofectamine 2000 in primary B cells. (PDF 324 kb)

Supplementary Methods (PDF 102 kb)

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Cai, D., Mataraza, J., Qin, ZH. et al. Highly efficient molecular delivery into mammalian cells using carbon nanotube spearing. Nat Methods 2, 449–454 (2005). https://doi.org/10.1038/nmeth761

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