The ISG15 Knockout 143B Polyclonal Cells are a CRISPR/Cas9-edited polyclonal population derived from the human 143B osteosarcoma cell line, designed for targeted disruption of ISG15 to generate a loss-of-function model. This product provides a heterogeneous knockout pool suitable for studying ISG15-dependent functions in a tumorigenic background.
The 143B cell line is derived from a primary human osteosarcoma, harboring a mutant TP53 (p53) gene, and exhibits aggressive growth and high metastatic potential in mouse models. Widely employed as a model for osteogenic sarcoma, 143B cells are characterized by rapid proliferation, invasiveness, and impaired DNA repair, making them a stringent platform for cancer biology research.
ISG15 is an interferon-stimulated ubiquitin-like protein that conjugates to target lysines via the E1 enzyme UBE1L, E2 UbcH8, and E3 HERC5, forming ISGylation. Transcription is activated by type I IFNs through the JAK-STAT pathway, involving the ISGF3 complex (STAT1-STAT2-IRF9) and IRF3/IRF7. ISG15 modifies proteins such as IRF3, MDA5, RIG-I, STAT1, filamin B, and PKR, modulating antiviral signaling, protein stability, and cytoskeletal organization. DeISGylation is performed by USP18, and free ISG15 can be secreted to act on immune cells. Interactions with HDAC6 and p62 further connect ISG15 to autophagy.
In osteosarcoma, ISG15 may influence tumor progression and metastatic spread. The 143B line??s high metastatic ability, coupled with ISG15??s role in filamin B modification and interferon-driven signaling, makes this knockout model valuable for exploring how ISGylation affects cell migration, invasion, and in vivo tumorigenesis. It also provides a tool to examine crosstalk between innate immune pathways and bone cancer aggressiveness.
Applications include Western blotting for ISG15 and ISGylation substrates, RT-qPCR for transcriptional analysis, interferon stimulation assays, proliferation (MTT/BrdU), migration/invasion transwell assays, and tumor xenograft studies. Flow cytometry can profile immune markers, and co-culture experiments with NK cells or T cells may assess paracrine cytokine effects. These polyclonal knockout cells are also suited for drug response modifier screens. For additional information, please contact Ascent Research.
