The KCNC4 Knockout HEK293T Polyclonal Cells product provides a CRISPR/Cas9-edited polyclonal knockout cell population in which the KCNC4 gene has been disrupted, leading to abrogation of the Kv3.4 voltage-gated potassium channel. This polyclonal pool, generated from the highly transfectable HEK293T line, offers a genetically diverse, constitutive knockout model that minimizes clone-specific artifacts and is ideal for reproducible electrophysiological and pharmacological assays.
HEK293T cells, a human embryonic kidney derivative, stably express the SV40 large T antigen, allowing episomal replication of SV40 origin-containing plasmids and resulting in exceptionally high transfection efficiencies. They are extensively used for recombinant protein expression, lentiviral production, and genome engineering applications. The absence of endogenous voltage-gated potassium conductances makes them an ideal host for heterologous KCNC4 expression and knockout studies, ensuring that measured currents originate solely from the introduced channel.
KCNC4 encodes Kv3.4, a Shaw-related voltage-gated potassium channel subunit with rapid activation kinetics. Depolarization opens the channel via its voltage sensor domain, permitting selective K+ efflux through the pore domain, which drives membrane repolarization and shortens action potential duration, thereby supporting high-frequency neuronal firing. Kv3.4 is modulated by protein kinase C (PKC)-mediated phosphorylation and by association with KCNE auxiliary subunits that alter gating and expression. It interacts with KCNC1 and KCNC2 to form heteromeric channels, and its activity influences neurotransmitter release by limiting Ca2+ influx. Disruption of KCNC4 eliminates this key repolarizing current, providing a clean background for studying channelopathies.
In the HEK293T heterologous system, KCNC4 knockout cells enable precise structure?Cfunction analysis of Kv3.4 without interference from native conductances. The polyclonal nature ensures that results reflect a population-wide knockout rather than a single clonal isolate, increasing experimental reproducibility. These cells are particularly valuable for validating antibody specificity and for high-throughput screening of channel modulators, as they offer a definitive negative control and a consistent cellular background for automated patch-clamp platforms. They also facilitate investigation of channel regulation by PKC and KCNE subunits when used in complementation assays with wild-type or mutant KCNC4 constructs.
Key applications include whole-cell and single-channel patch-clamp electrophysiology to characterize Kv3.4 biophysics and pharmacology, high-throughput screening for agonists and antagonists, and molecular analyses via western blotting, immunofluorescence, and RT-qPCR to confirm knockout and assess expression. The cells are suited for modeling potassium channelopathies linked to epilepsy, neurodevelopmental disorders, and cardiac arrhythmias. For technical inquiries, custom experimental design, or further details, please contact Ascent Research.
