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

Methods Mol Biol
Authors
Keywords
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
Journal
Methods Mol Biol
Volume
1787
Pages
235-252
Date Published
2018
ISSN
1940-6029
DOI
10.1007/978-1-4939-7847-2_18
PubMed ID
29736723
Links
Grant list
R01 MH115045 / MH / NIMH NIH HHS / United States