The C12orf57 Knockout SK-HEP-1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout population derived from SK-HEP-1 cells, with targeted disruption of the C12orf57 gene. This mixed knockout pool provides a loss-of-function model suitable for functional genomics and disease modeling, avoiding clonal selection biases. The polyclonal format ensures broad applicability in studying gene function within a heterogeneous cell population.
SK-HEP-1 is an immortalized human hepatic adenocarcinoma cell line, adherent and epithelial-like, derived from a 52-year-old male patient. Originally misclassified as endothelial, it is now confirmed as an epithelial adenocarcinoma, serving as a widely used model for liver cancer research including tumor invasiveness and drug resistance studies.
C12orf57 encodes a protein essential for the O-mannosylation of dystroglycan, a process critical for cell-matrix adhesion. It interacts with glycosyltransferases POMT1, POMT2, and B3GALNT2, and with FKTN and FKRP, to facilitate the assembly of the functional glycan chain on alpha-dystroglycan. This modification enables dystroglycan to bind laminin with high affinity, thereby linking the extracellular matrix to the cytoskeleton. Loss of C12orf57 results in dystroglycan hypoglycosylation, impairing adhesion and downstream integrin-mediated signals. Additionally, C12orf57 modulates apoptosis through BCL2 family members, positioning it at a regulatory node between cell survival and biomechanical sensing. Together with B4GAT1 and LARGE1, these components constitute the core machinery of dystroglycan glycosylation.
The C12orf57 knockout in SK-HEP-1 cells offers a clinically relevant model to study dystroglycanopathy in liver cancer. Temtamy syndrome, linked to C12orf57 mutations, includes neurodevelopmental defects, but its impact on hepatic tissue is poorly understood. This polyclonal knockout model facilitates the investigation of how dystroglycan hypoglycosylation influences hepatocellular carcinoma cell adhesion, migration, and apoptotic resistance, potentially revealing new therapeutic targets. It also enables examination of tumor microenvironment interactions and metastatic behavior in the context of glycosylation defects.
Researchers can utilize this model for western blotting and lectin blotting to assess dystroglycan glycosylation, immunofluorescence for dystroglycan localization, flow cytometry for adhesion molecule profiling, and functional assays such as migration, invasion, and apoptosis detection. RT-qPCR enables analysis of glycosylation gene expression. Applications include investigating C12orf57 in hepatocellular carcinoma, modeling Temtamy syndrome pathology, drug sensitivity screening, and studying O-mannosylation in liver cells. For further assistance, contact Ascent Research.
