The KCNJ14 Knockout HEK293T Polyclonal Cells constitute a CRISPR/Cas9-mediated gene disruption model designed to abolish expression of the human KCNJ14 gene, which encodes the inward rectifier potassium channel Kir2.4. This product is supplied as a polyclonal cell population, reflecting a heterogeneous pool of edited cells generated via transient delivery of CRISPR/Cas9 reagents rather than a clonally derived line. The resulting loss-of-function model enables systematic investigation of KCNJ14-dependent signaling and electrophysiological properties in a readily transfectable human cell background.
HEK293T cells are an extensively characterized human embryonic kidney epithelial line immortalized by stable expression of the SV40 large T antigen and transformed with adenovirus 5 DNA. Their high transfection efficiency and robust protein expression capacity make them a universal workhorse for mammalian cell studies. In the context of genome editing, these features facilitate efficient delivery of CRISPR components and subsequent selection for edited pools, yielding a versatile polyclonal knockout population that retains the core characteristics of the parental line while eliminating target gene function across a spectrum of genetic lesions.
KCNJ14 encodes the inward rectifier potassium channel Kir2.4, which stabilizes the resting membrane potential and dampens excitability by facilitating potassium efflux at hyperpolarized potentials. Channel activity is regulated by phosphatidylinositol 4,5-bisphosphate (PIP2), protein kinase A (PKA), protein kinase C (PKC), G protein ?¦? subunits, intracellular Mg2+, and pH, integrating G protein-coupled receptor (GPCR) signaling with electrical properties. GPCR activation modulates adenylate cyclase and cAMP levels, altering PKA-mediated phosphorylation of Kir2.4; PIP2 dynamics, controlled by phospholipase C, directly affect gating. Downstream, Kir2.4 influences voltage-gated calcium channel function and neuronal excitability. Scaffolding proteins PSD-95 and SAP97 facilitate its subcellular localization, and heteromeric assembly with Kir2.1, Kir2.2, and Kir2.3 further diversifies its physiological roles.
In the HEK293T cellular context, this polyclonal knockout model offers a robust platform for dissecting the contribution of KCNJ14 to membrane potential control and GPCR signal integration, even though HEK293T cells are not classically excitable. The parental line expresses many signaling components, including GPCRs, adenylate cyclase, and PKA, enabling reconstitution of regulatory networks upon exogenous expression of relevant receptors. The polyclonal nature of the knockout population mitigates clonal bias and allows assessment of gene function in a genetically diverse background, better approximating the heterogeneity found in tissue or disease models. Consequently, this product is particularly suited for high-throughput screening of channel modulators where population-level responses are more representative than those from single-cell clones.
Researchers can apply these KCNJ14 knockout HEK293T cells in patch-clamp electrophysiology, calcium imaging, and cell viability assays to probe channel-dependent phenomena. The polyclonal population is ideal for functional genomics, drug screening for channel modulators, and modeling potassium channelopathies associated with retinal degeneration and neurological disorders. Standard assays such as Western blotting, RT-qPCR, and immunofluorescence confirm KCNJ14 depletion. For further information or inquiries, please contact Ascent Research.
