SLC12A6 Knockout Cell Line (HEK293)

The choice depends on whether you are studying SLC12A6 (KCC3)'s role as a K⁺-Cl⁻ cotransporter or modeling Andermann syndrome (agenesis of corpus callosum with peripheral neuropathy). The Knockout line is the standard tool for asking whether KCC3 is required for K⁺-Cl⁻ efflux — KCC3 is broadly expressed and contributes to cell volume regulation, ion homeostasis, and neuronal chloride extrusion. Overexpression is useful for studying transport activity or for testing disease-associated mutations. For KCC family research, the EDITGENE SLC12A6 Knockout in HEK293 is the standard mechanistic platform — HEK293 has been extensively used for KCC family biochemistry. SLC12A6 mutations cause Andermann syndrome (autosomal recessive ACCPN) — disease variant rescue enables genotype-function studies. Rescue with wild-type, transport-deficient, or phospho-mimetic (constitutively active, T991A/T1048A) KCC3 enables comprehensive structure-function and regulatory mechanism studies.

Primary applications: • K⁺-Cl⁻ cotransport: cellular K⁺ and Cl⁻ flux measurements using radiotracer or fluorescent probe-based assays. • Cell volume regulation: regulatory volume decrease (RVD) assays following hypotonic challenge given KCC3's role in cell volume control. • Andermann syndrome modeling: rescue with disease-associated SLC12A6 mutations for genotype-function correlation studies. • Regulatory mechanism studies: WNK kinase-SPAK/OSR1 pathway analysis given KCC3 regulation through phosphorylation of conserved threonines (T991, T1048). EDITGENE recommends this model for researchers investigating K⁺-Cl⁻ cotransport biology, cell volume regulation, and Andermann syndrome mechanisms.

Yes. KCC3 rescue experiments are well-established for cation-chloride cotransporter research: • Construct design: use a codon-modified SLC12A6 sequence with a small C-terminal tag (FLAG, HA). KCC3 has 12 transmembrane domains with a large cytoplasmic C-terminus. • Phospho-regulatory rescue: T991A/T1048A double mutation creates constitutively active KCC3, useful for distinguishing transport from regulatory effects. • Andermann syndrome rescue: patient-derived SLC12A6 mutations enable disease genotype-function correlation. • Functional readout: rescue should restore K⁺-Cl⁻ cotransport activity measured by ⁸⁶Rb⁺ flux or cell volume regulatory volume decrease. HEK293 transduces efficiently with lentivirus and supports stable rescue line generation.