|Publication Type||Journal Article|
|Year of Publication||2018|
|Authors||Pan, JQ, Baez-Nieto, D, Allen, A, Wang, H-R, Cottrell, JR|
|Journal||Methods Mol Biol|
|Keywords||Calcium Channels, Data Interpretation, Statistical, Drug Discovery, Electrophysiological Phenomena, Gene Expression, HEK293 Cells, High-Throughput Screening Assays, Humans, Ion Channel Gating, Patch-Clamp Techniques, Workflow|
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.
|Alternate Journal||Methods Mol Biol|
|Grant List||R01 MH115045 / MH / NIMH NIH HHS / United States|