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Nonviral ultrasound-mediated gene delivery in small and large animal models

Nature Protocols volume 14, pages 1015–1026 (2019)Cite this article

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Abstract

Ultrasound-mediated gene delivery (sonoporation) is a minimally invasive, nonviral and clinically translatable method of gene therapy. This method offers a favorable safety profile over that of viral vectors and is less invasive as compared with other physical gene delivery approaches (e.g., electroporation). We have previously used sonoporation to overexpress transgenes in different skeletal tissues in order to induce tissue regeneration. Here, we provide a protocol that could easily be adapted to address various other targets of tissue regeneration or additional applications, such as cancer and neurodegenerative diseases. This protocol describes how to prepare, conduct and optimize ultrasound-mediated gene delivery in both a murine and a porcine animal model. The protocol includes the preparation of a microbubble–DNA mix and in vivo sonoporation under ultrasound imaging. Ultrasound-mediated gene delivery can be accomplished within 10 min. After DNA delivery, animals can be followed to monitor gene expression, protein secretion and other transgene-specific outcomes, including tissue regeneration. This procedure can be accomplished by a competent graduate student or technician with prior experience in ultrasound imaging or in performing in vivo procedures.

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Fig. 1: Spacer production.
Fig. 2: Sonoporation in mice.
Fig. 3: Sonoporation in pigs.
Fig. 4: Anticipated results.

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Data availability

All materials are available from commercial sources or can be derived using methods described in this study. All relevant data are reported in the article.

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Acknowledgements

We acknowledge funding support from the California Institute for Regenerative Medicine (CIRM; (grant TR4-06713 to D.G.) and NIH grants R01EB026094 (to D.G.) and R01CA112356 (to K.W.F.).

Author information

Authors and Affiliations

  1. Skeletal Biotech Laboratory, The Hebrew University–Hadassah Faculty of Dental Medicine, Ein Kerem, Jerusalem, Israel

    Maxim Bez, Galina Shapiro, Gadi Pelled & Dan Gazit

  2. Department of Surgery, Cedars-Sinai Medical Center, Los Angeles, CA, USA

    Maxim Bez, Gadi Pelled & Dan Gazit

  3. Department of Radiology, Stanford University School of Medicine, Stanford, CA, USA

    Josquin Foiret & Katherine W. Ferrara

  4. Board of Governors Regenerative Medicine Institute, Cedars-Sinai Medical Center, Los Angeles, CA, USA

    Gadi Pelled & Dan Gazit

  5. Department of Biomedical Sciences, Cedars-Sinai Medical Center, Los Angeles, CA, USA

    Gadi Pelled & Dan Gazit

  6. Biomedical Imaging Research Institute, Cedars-Sinai Medical Center, Los Angeles, CA, USA

    Gadi Pelled & Dan Gazit

  7. Department of Orthopedics, Cedars-Sinai Medical Center, Los Angeles, CA, USA

    Dan Gazit

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  1. Maxim Bez
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Contributions

G.P., K.W.F. and D.G. designed the experiments. M.B., J.F. and G.S. performed the experiments. M.B. and G.S. analyzed the data. M.B., J.F. and G.S. created the figures. M.B., J.F. and G.S. wrote the manuscript. All authors approved the manuscript.

Corresponding author

Correspondence to Dan Gazit.

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Competing interests

G.P. and D.G. are shareholders in GamlaStem Medical, which did not provide funds for this study. The other authors declare no competing interests.

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Key references using this protocol

Shapiro, G. et al. J. Control. Release 223, 157–164 (2016): https://doi.org/10.1016/j.jconrel.2015.12.001

Bez, M. et al. Sci. Transl. Med. 9, eaal3128 (2017): https://doi.org/10.1126/scitranslmed.aal3128

Bez, M. et al. Mol. Ther. 26, 1746–1755 (2018): https://doi.org/10.1016/j.ymthe.2018.04.020

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Bez, M., Foiret, J., Shapiro, G. et al. Nonviral ultrasound-mediated gene delivery in small and large animal models. Nat Protoc 14, 1015–1026 (2019). https://doi.org/10.1038/s41596-019-0125-y

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