The ISG15 Knockout 769-P Polyclonal Cells product provides a CRISPR/Cas9-edited polyclonal knockout cell population in which the ISG15 gene is disrupted within the 769-P human clear cell renal cell carcinoma (ccRCC) host line. This polyclonal format maintains a heterogeneous pool of knockout alleles, enabling population-level loss-of-function analyses while minimizing clonal artifacts. The cell pool is derived from a well-characterized primary ccRCC isolate, offering a relevant genetic background for investigating ISG15-dependent innate immune signaling, interferon responses, and tumor biology without the confounding effects of clonal selection.
The 769-P cell line was established from a primary clear cell renal cell carcinoma and serves as a clinically relevant in vitro model for ccRCC, a kidney cancer subtype characterized by dysregulated hypoxia and immune signaling pathways. These cells retain key molecular features of renal carcinoma, including aberrant interferon response signatures and constitutive activation of JAK-STAT signaling components. The 769-P background is extensively employed to dissect tumor-intrinsic innate immune programs, viral susceptibility, and response to immunomodulatory agents, making it an ideal host for ISG15 knockout studies.
ISG15 encodes a ubiquitin-like protein strongly induced by type I interferons (IFN-??/??) via ISRE elements bound by IRF3, IRF7, and STAT1/STAT2/IRF9 complexes. Post-translationally, ISG15 is conjugated to target lysines through an enzymatic cascade involving E1 UBE1L, E2 UBCH8 (UBE2L6), and E3 ligases HERC5/HERC6; USP18 reverses this ISGylation. Key substrates include JAK1, STAT1, IRF3, PKR, IFIT proteins, and OAS proteins, and ISGylation modulates their stability, localization, and activity. Consequently, ISG15 amplifies JAK-STAT signaling, promotes interferon-stimulated gene expression, and establishes an antiviral cellular state.
In the 769-P clear cell renal carcinoma context, ISG15 knockout abrogates ISGylation and alters interferon signaling dynamics. ccRCC tumors often show dysregulated JAK-STAT and interferon signatures; this model distinguishes whether ISG15 acts as a tumor suppressor by enhancing antiviral immunity and inhibiting growth, or supports oncogenic pathways via survival signaling and immune evasion. Researchers can evaluate how ISG15 loss affects proliferation, invasion, interferon therapy sensitivity, and oncolytic virus response, clarifying innate immunity in renal cancer biology.
This knockout cell pool supports diverse experimental applications. Interferon signaling studies employ IFN-??/??/?? stimulation with RT-qPCR and western blotting for STAT phosphorylation and ISG expression. Antiviral assays involve RNA or DNA virus infection and replication monitoring. Cancer-relevant readouts include proliferation, migration, and invasion assays under basal and interferon-treated conditions. Co-immunoprecipitation and proteomics facilitate ISGylation substrate discovery, and drug response profiling plus flow cytometry enable tumor microenvironment modeling. For further information or custom inquiries, please contact Ascent Research.
