The JPH1 Knockout SK-HEP-1 Polyclonal Cells consist of a CRISPR/Cas9-edited polyclonal knockout population derived from the SK-HEP-1 human liver adenocarcinoma cell line, featuring targeted disruption of the JPH1 gene. This heterogeneous pool is not a monoclonal line, providing a versatile loss-of-function model that avoids single-cell clonal selection and enables pooled functional studies of junctophilin-1 at endogenous expression levels.
The parental SK-HEP-1 cell line was originally isolated from the ascites of a patient with liver adenocarcinoma and exhibits non-parenchymal, endothelial-like characteristics, co-expressing mesenchymal and endothelial markers. This unique phenotype makes SK-HEP-1 a valuable host for modeling hepatic adenocarcinoma and endothelial-to-mesenchymal transition, allowing researchers to examine how JPH1 loss impacts cancer cell adhesion, migration, and calcium-dependent signaling pathways in a liver cancer context.
Junctophilin-1 (JPH1) is a structural protein that anchors the endoplasmic reticulum (ER) to the plasma membrane via its membrane occupation and recognition nexus (MORN) domains, creating junctional complexes essential for localized calcium signaling. Within these junctions, JPH1 directly interacts with ER calcium release channels including inositol 1,4,5-trisphosphate receptors (IP3Rs) and ryanodine receptors (RyR1/RyR2), and forms complexes with SERCA2, caveolin-1, and calmodulin to govern calcium microdomain dynamics. JPH1 expression is transcriptionally regulated by MEF2, PGC-1??, and the calcium/calcineurin/NFAT pathway; downstream, JPH1-mediated calcium signaling activates NFAT transcriptional programs and CaMKII phosphorylation, coupling extracellular stimuli to efficient intracellular calcium responses.
Disruption of JPH1 in SK-HEP-1 cells is expected to perturb ER-PM junctional architecture and decouple localized calcium release from downstream effectors. Given the emerging role of calcium dysregulation in hepatocellular carcinoma progression, this polyclonal knockout model facilitates interrogation of JPH1 function in cancer cell proliferation, migration, and survival. The endothelial-like features of SK-HEP-1 further permit investigation of junctional complex contributions to cell motility and angiogenic signaling, offering a multifaceted tool for mechanistic cancer studies.
Key experimental applications include calcium imaging with Fluo-4 or GCaMP indicators, co-immunoprecipitation for mapping JPH1 protein interactors, and qRT-PCR or western blotting to verify target ablation and downstream signaling changes. Phenotypic assays such as MTT proliferation, Transwell migration, and flow cytometry?Cbased calcium flux measurements are well-suited for functional characterization. RNA-sequencing of the knockout pool can reveal transcriptome-wide consequences of disrupted ER-PM tethering. For detailed product information or technical support, please contact Ascent Research.
