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Helios® Gene Gun–Mediated Transfection of the Inner Ear Sensory Epithelium

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Auditory and Vestibular Research

Part of the book series: Methods in Molecular Biology™ ((MIMB,volume 493))

Helios® Gene Gun–mediated transfection is a biolistic method for mechanical delivery of exogenous DNA into cells in vitro or in vivo. The technique is based on bombardment of a targeted cellular surface by micron- or submicron-sized DNA-coated gold particles that are accelerated by a pressure pulse of compressed helium gas. The main advantage of Helios® Gene Gun–mediated transfections is that it functions well on various cell types, including terminally differentiated cells that are difficult to transfect, such as neurons or inner ear sensory hair cells, and cells in internal cellular layers, such as neurons in organotypic brain slices. The successful delivery of mRNA, siRNA, or DNA of practically any size can be achieved using biolistic transfection. This chapter provides a detailed description and critical evaluation of the methodology used to transfect cDNA expression constructs, including green fluorescent protein (GFP) tagged full-length cDNAs of myosin XVa, whirlin, and β-actin, into cultured inner ear sensory epithelia using the Bio-Rad Helios® Gene Gun.

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Acknowledgments

I thank Thomas Friedman for support and encouragement as well as for critical reading of this chapter, Jonathan Gale (University of London, UK) from whom I learned the organ of Corti explant technique, Erich Boger for preparing myosin XVa and whirlin cDNA expression constructs, critical reading of the chapter and helpful discussions, Andrew Griffith, Valentina Labay, Polina Belyantseva and Shin-Ichiro Kitajiri for critical reading of the manuscript. Supported by funds from the NIDCD Intramural Program (1 Z01 DC 00035-11) to Thomas B. Friedman.

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Authors and Affiliations

  1. Laboratory of Molecular Genetics, Section on Human Genetics, NIDCD, National Institutes of Health, Rockville MD, USA

    Inna A. Belyantseva

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  1. Inna A. Belyantseva
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Editors and Affiliations

  1. University of South Florida, College of Medicine, Tampa, FL, USA

    Bernd Sokolowski

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© 2009 Humana Press, a part of Springer Science+Business Media, LLC

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Belyantseva, I.A. (2009). Helios® Gene Gun–Mediated Transfection of the Inner Ear Sensory Epithelium. In: Sokolowski, B. (eds) Auditory and Vestibular Research. Methods in Molecular Biology™, vol 493. Humana Press. https://doi.org/10.1007/978-1-59745-523-7_7

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  • DOI: https://doi.org/10.1007/978-1-59745-523-7_7

  • Publisher Name: Humana Press

  • Print ISBN: 978-1-934115-62-6

  • Online ISBN: 978-1-59745-523-7

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