Description

The ANK3 Knockout HK-2 Cell Line is a CRISPR/Cas9-edited knockout cell line derived from the HK-2 human proximal tubule epithelial cell line. This loss-of-function model enables targeted disruption of the ANK3 gene, which encodes ankyrin-G, a key adapter protein linking integral membrane proteins to the spectrin-actin cytoskeleton. This cell line provides a genetically defined system for investigating ankyrin-G-dependent processes in renal epithelial biology.

The HK-2 cell line is an immortalized human kidney proximal tubule epithelial cell line derived from normal adult kidney. It retains many characteristics of primary proximal tubule cells, including polarized morphology, expression of brush border enzymes, and capacity for solute reabsorption, secretion, and metabolic clearance. HK-2 cells are widely used as a model for renal drug transport, nephrotoxicity, and epithelial barrier function studies.

ANK3 encodes ankyrin-G, which anchors integral membrane proteins such as voltage-gated sodium and potassium channels, E-cadherin, and neurofascin to the spectrin-actin network. This anchoring is essential for ion channel clustering, maintenance of cell adhesion complexes, and signal transduction. Ankyrin-G functions downstream of Wnt3a and TGF-?? signaling, interacting with ??-catenin and E-cadherin to modulate Wnt/??-catenin pathway activity. It also participates in PI3K-Akt signaling. Disruption of ANK3 impairs these interactions, leading to compromised cytoskeletal integrity and altered downstream effector activation, including NF-??B.

In the HK-2 proximal tubule context, ANK3 knockout disrupts ankyrin-G-mediated assembly of tight junctions and ion channel complexes, thereby impairing epithelial barrier function and vectorial transport. This model recapitulates aspects of renal tubular dysfunction and is relevant to research on electrolyte imbalance, drug-induced nephrotoxicity, and the renal manifestations of disorders linked to ANK3, such as bipolar disorder and cardiac arrhythmia. The HK-2 background provides a physiologically relevant platform to study how loss of ankyrin-G affects proximal tubule homeostasis.

This knockout cell line supports diverse experimental applications, including kidney disease modeling, nephrotoxicity screening, and mechanistic investigation of solute transport. Researchers can employ techniques such as transepithelial electrical resistance (TEER) measurement, immunofluorescence, western blotting, RT-qPCR, and dual-luciferase Wnt reporter assays. The model is also suitable for co-immunoprecipitation studies of cytoskeletal and adhesion complexes, migration assays, and flow cytometry. For further details or to inquire about custom services, please contact Ascent Research.