TMEM63A Knockout Hela Cell Line

The TMEM63A Knockout HeLa Cell Line is a CRISPR/Cas9-edited knockout cell line with targeted disruption of the TMEM63A gene in the HeLa background. This loss-of-function model eliminates endogenous TMEM63A expression, providing a robust system for studying the mechanosensitive cation channel in an epithelial context. It enables precise functional analysis of calcium-dependent mechanotransduction and osmoregulation.

HeLa is an immortalized human cervical adenocarcinoma cell line, derived from Henrietta Lacks in 1951. Widely used in cancer biology and epithelial research, its epithelial origin and well-characterized signaling networks make it an ideal host for investigating mechanosensitive ion channels. The HeLa background facilitates studies of cell volume regulation and mechanical stress responses in a neoplastic environment.

TMEM63A encodes a calcium-permeable channel activated by hypo-osmotic stress, mechanical stretch, and cell swelling. Channel opening leads to calcium influx, which triggers downstream effectors including calmodulin, calcineurin, and CAMK, ultimately regulating NFAT-dependent transcription. Thus, TMEM63A functions upstream of calcium-dependent signaling cascades. TMEM63A interacts with cytoskeletal components, coupling mechanical cues to calcium signaling and cell volume control. This pathway is central to mechanotransduction and osmotic adaptation.

In HeLa cells, TMEM63A knockout allows dissection of epithelial mechanosensitive signaling. Loss of TMEM63A can reveal its specific contributions to calcium dynamics, cytoskeletal remodeling, and volume regulation under mechanical stimuli. The model is relevant to neurological disorders such as hypomyelinating leukodystrophy, where TMEM63A mutations are implicated, offering a platform for mechanistic studies in a human cell system.

Applications include calcium imaging with Fluo-4 upon hypo-osmotic shock, patch-clamp electrophysiology, cell volume regulation assays, and screening for mechanosensitive channel modulators. This cell line also supports western blotting and immunofluorescence for TMEM63A validation, and serves as a clean host for expressing TMEM63A mutants to study structure-function relationships. For additional product information, please contact Ascent Research.