Protocol to study human Kv1.2 potassium channel pathogenic sequence variants using two-electrode voltage-clamp technique
Year: 2025
Type: article
Abstract: The abietane diterpenoid pisiferic acid (PA) from conifer Chamaecyparis pisifera is a pan-rescuer of human Kv1.2 channel pathogenic loss-of-function (LOF) sequence variants. Here, we present a protocol for the study of Kv1.2 channel sequence variants using the Xenopus laevis oocyte expression system and two-electrode voltage-clamp (TEVC) electrophysiology. We describe steps for studying mutant Kv1.2, wild-type Kv1.2, and wild-type Kv1.1 cRNA combinations in Xenopus laevis oocytes; performing TEVC; and data analysis. For complete details on the use and execution of this protocol, please refer to Manville et al. 1 • Instructions for cRNA preparation and expression of ion channels in Xenopus oocytes • Guide to recording currents from ion channels in Xenopus oocytes using TEVC • Steps for analyzing TEVC data recorded from ion channels expressed in Xenopus oocytes Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics. The abietane diterpenoid pisiferic acid (PA) from conifer Chamaecyparis pisifera is a pan-rescuer of human Kv1.2 channel pathogenic loss-of-function (LOF) sequence variants. Here, we present a protocol for the study of Kv1.2 channel sequence variants using the Xenopus laevis oocyte expression system and two-electrode voltage-clamp (TEVC) electrophysiology. We describe steps for studying mutant Kv1.2, wild-type Kv1.2, and wild-type Kv1.1 cRNA combinations in Xenopus laevis oocytes; performing TEVC; and data analysis.
Source: STAR Protocols
Authors: Rían W. Manville, Geoffrey W. Abbott
Institution: University of California, Irvine
Language: English
Subfield: Molecular Biology
Domain: Life Sciences
Open Access status: gold
Estimated APC paid: $1,550
OpenAlex ID: W4416538848