The CAPN15 Knockout HeLa Cell Line is a CRISPR/Cas9-edited knockout cell line engineered from the HeLa host. This model features targeted disruption of the CAPN15 gene, encoding a calcium-dependent cysteine protease. It serves as a loss-of-function tool to dissect CAPN15-mediated proteolytic events, created via CRISPR/Cas9 to ensure stable gene disruption without transgene insertion.
HeLa is a cervical adenocarcinoma epithelial line positive for HPV18. The viral oncoproteins E6 and E7 inhibit p53 and Rb, respectively, resulting in a highly proliferative and aneuploid phenotype. Derived from a cervical cancer biopsy, HeLa cells are a cornerstone of cancer research, virology, and cell biology, and provide a robust host for CRISPR/Cas9-mediated knockout studies. However, their transformed nature should be taken into account when interpreting CAPN15-dependent effects.
CAPN15 is a calcium-activated cysteine protease that carries out limited proteolysis of key substrates, including spectrin, talin, the apoptosis regulators Bax and Bid, and various signaling kinases. Its activity is induced by calcium influx and integrin-mediated signaling, and is tightly controlled by the endogenous inhibitor calpastatin (CAST). CAPN15 interacts with calcium-binding proteins and integrin complexes, and functions within a network that includes calmodulin (CALM1) and calcium channels. Through its proteolytic action, CAPN15 modulates apoptosis, cell migration, and cytoskeletal dynamics.
In HeLa cells, where p53 and Rb pathways are already suppressed by HPV oncoproteins, CAPN15 knockout provides a powerful system to study calcium-regulated proteolysis in a cancer context. Disruption of CAPN15 may impair apoptotic signaling, alter cytoskeletal integrity, and reduce migratory capacity??all processes implicated in tumor progression. This model is particularly relevant for cervical adenocarcinoma research and offers a platform to investigate calpain-related mechanisms in neurodegenerative diseases and cataracts. It enables precise analysis of substrate turnover and cellular responses to calcium fluctuations.
Typical applications include apoptosis detection with Annexin V/PI staining, cell migration assessment using transwell or scratch assays, and immunofluorescence visualization of cytoskeletal proteins such as spectrin or talin. Western blotting confirms CAPN15 knockout and downstream target cleavage, while calpain activity assays provide functional validation. This cell line is also suited for drug target screening of calpain inhibitors. Side-by-side comparison with wild-type HeLa allows dissection of CAPN15-specific functions. For further details, please contact Ascent Research.
