Developing High-Throughput Assays to Analyze and Screen Electrophysiological Phenotypes.

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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.

Full Text

Duke Authors

Cited Authors

  • Pan, JQ; Baez-Nieto, D; Allen, A; Wang, H-R; Cottrell, JR

Published Date

  • 2018

Volume / Issue

  • 1787 /

Start / End Page

  • 235 - 252

PubMed ID

  • 29736723

Pubmed Central ID

  • 29736723

Digital Object Identifier (DOI)

  • 10.1007/978-1-4939-7847-2_18