2018 - Developing High-Throughput Assays to Analyze and Screen Electrophysiological Phenotypes

icon sp96   SyncroPatch 384PE book chapter in Phenotypic Screening (2018)

Authors:
Pan J.Q., Baez-Nieto D., Allen A., Wang HR., Cottrell J.R.

Book Chapter:
In: Wagner B. (eds) Phenotypic Screening. Methods in Molecular Biology, vol 1787. Humana Press, New York, NY


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.


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