Developing High-Throughput Assays to Analyze and Screen Electrophysiological Phenotypes.
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Abstract | Ion channels represent nearly a quarter of all targets that currently available medications modulate, and their dysfunction underlies increasing number of human diseases. Functional analysis of ion channels have traditionally been a bottleneck in large-scale analyses. Recent technological breakthroughs in automated planar electrophysiology have democratized the technique to enable high-throughput patch clamping at scale. In this chapter, we describe the methodology to perform a phenotypic screen on voltage-gated calcium channels across many different genetic coding variations and against small-molecule modulators. We first describe the procedures to establish inducible heterologous ion channel expression in HEK293 cells, where each cell incorporates one copy of a target protein cDNA-a step that is critical for producing stable and consistent expression of ion channels. We then describe the experimental and analytical methods for analyzing the function of ion channels using high-throughput planar electrophysiology. |
Year of Publication | 2018
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Journal | Methods Mol Biol
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Volume | 1787
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Pages | 235-252
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Date Published | 2018
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ISSN | 1940-6029
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DOI | 10.1007/978-1-4939-7847-2_18
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PubMed ID | 29736723
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Grant list | R01 MH115045 / MH / NIMH NIH HHS / United States
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