The CD63 Knockout 786-O Polyclonal Cells constitute a CRISPR/Cas9-edited polyclonal knockout cell population derived from the 786-O human renal cell carcinoma line, engineered to disrupt the CD63 gene. This product provides a heterogeneous pool of cells carrying diverse loss-of-function alleles, enabling robust interrogation of CD63-dependent processes without the clonal bias inherent in monoclonal lines. The polyclonal format is well-suited for population-level functional assays, supporting the analysis of exosome-related pathways, adhesion dynamics, and migratory behavior in a genetically diverse knockout background. Researchers can apply standard selection and screening protocols to validate target-gene disruption, leveraging the broad applicability of this model in advanced cell biology and cancer research workflows.
Derived from a primary clear cell adenocarcinoma of the kidney, 786-O cells serve as a flagship model for clear cell renal cell carcinoma (ccRCC). The line is VHL-null, leading to constitutive stabilization of hypoxia-inducible factors (HIFs) and pseudohypoxic signaling, recapitulating the hypervascular and metabolically reprogrammed phenotype characteristic of ccRCC. 786-O cells exhibit robust tumorigenic capacity in xenograft models and are commonly employed to study VEGF-driven angiogenesis, HIF-mediated transcription, and metabolic adaptations. This well-characterized genetic background provides a clinically relevant context for assessing the contribution of CD63 to renal cancer progression and lysosome-related organelle function.
CD63 encodes a tetraspanin protein that orchestrates the formation of tetraspanin-enriched microdomains, which serve as platforms for exosome biogenesis and cargo sorting through interactions with syntenin-1 and the ESCRT accessory protein ALIX. CD63 also engages integrin ??1, CD9, CD81, and CD151 to regulate focal adhesion dynamics and cell migration, downstream of regulators such as hypoxia, MITF, and cellular stress signals. It transduces signals via integrin-mediated cascades (PI3K/AKT and ERK) and influences exosomal miRNA transfer, matrix metalloproteinase activity, and adhesion complex remodeling. The tetraspanin web, including partners like Rab27 GTPases and ESCRT-III components, ensures proper trafficking of adhesion receptors and multivesicular body maturation, highlighting CD63 as a nodal point linking membrane organization, secretion, and motility.
In the 786-O VHL-null setting, CD63 knockout disrupts a critical nexus between pseudohypoxic signaling and exosomal communication, potentially altering the dissemination of oncogenic cargo from ccRCC cells. Loss of CD63 is expected to impair integrin ??1 recycling and focal adhesion turnover, thereby attenuating cellular migration and invasion??processes central to metastasis. Concurrently, defective exosome secretion may modulate intercellular signaling within the tumor microenvironment, affecting stromal recruitment and immune evasion. This model thus enables dissection of how tetraspanin-dependent membrane dynamics intersect with HIF-driven pathways to control renal cancer cell behavior, aiding the identification of vulnerabilities in metastatic ccRCC.
Typical research applications include exosome biology studies, renal cell carcinoma metastasis modeling, lysosomal dysfunction investigations (e.g., Hermansky-Pudlak syndrome-related processes), and exploration of tetraspanin function in cell adhesion. Representative assays encompass CD63 immunoblotting, exosome isolation and proteomic profiling, transwell migration/invasion assays, integrin-based adhesion measurements, and flow cytometry for surface tetraspanin expression. The polyclonal knockout population also serves as a valuable comparative tool for rescue experiments, CRISPR screens, and pharmacological perturbation studies. For additional product specifications or technical consultation, please contact Ascent Research.
