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Nat Protoc. 2014 Oct;9(10):2438-49. doi: 10.1038/nprot.2014.168. Epub 2014 Sep 18.

Using injectoporation to deliver genes to mechanosensory hair cells.

Author information

1
1] Department of Molecular and Cellular Neuroscience, The Dorris Neuroscience Center, The Scripps Research Institute, La Jolla, California, USA. [2].
2
Department of Molecular and Cellular Neuroscience, The Dorris Neuroscience Center, The Scripps Research Institute, La Jolla, California, USA.

Abstract

Mechanosensation, the transduction of mechanical force into electrochemical signals, allows organisms to detect touch and sound, to register movement and gravity, and to sense changes in cell volume and shape. The hair cells of the mammalian inner ear are the mechanosensors for the detection of sound and head movement. The analysis of gene function in hair cells has been hampered by the lack of an efficient gene transfer method. Here we describe a method termed injectoporation that combines tissue microinjection with electroporation to express cDNAs and shRNAs in mouse cochlear hair cells. Injectoporation allows for gene transfer into dozens of hair cells, and it is compatible with the analysis of hair cell function using imaging approaches and electrophysiology. Tissue dissection and injectoporation can be carried out within a few hours, and the tissue can be cultured for days for subsequent functional analyses.

PMID:
25232939
PMCID:
PMC4241755
DOI:
10.1038/nprot.2014.168
[Indexed for MEDLINE]
Free PMC Article

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