Golden Gate cloning enables efficient concatemer construction for biophysical analysis of heterozygous potassium channel variants from patients with epilepsy

Missense mutations that cause channelopathies usually occur in a heterozygous setting. Functional voltage-gated potassium channels are tetramers of pore-forming α-subunits. When a variant is co-expressed with the wild-type gene, six distinct tetramers can assemble. Conventional techniques cannot be used to study the tetramers individually because they rely on simultaneous expression of separate genes resulting in a mixture of all possible …