Deep Mutational Scan of an SCN5A Voltage Sensor.

ABSTRACT:

Background

Variants in ion channel genes have classically been studied in low throughput by patch clamping. Deep mutational scanning is a complementary approach that can simultaneously assess function of thousands of variants.

Methods

We have developed and validated a method to perform a deep mutational scan of variants in SCN5A, which encodes the major voltage-gated sodium channel in the heart. We created a library of nearly all possible variants in a 36 base region of SCN5A in the S4 voltage sensor of domain IV and stably integrated the library into HEK293T cells.

Results

In preliminary experiments, challenge with 3 drugs (veratridine, brevetoxin, and ouabain) could discriminate wild-type channels from gain- and loss-of-function pathogenic variants. Hi

SUBMITTER: Glazer AM 

PROVIDER: S-EPMC7031040 | biostudies-literature | 2020 Feb

REPOSITORIES: biostudies-literature