The EFCAB14 Knockout 786-O Polyclonal Cells product provides a CRISPR/Cas9-edited polyclonal knockout population for studying the putative calcium sensor EFCAB14 in human renal cell adenocarcinoma. Engineered from the 786-O host cell line, this polyclonal knockout model enables loss-of-function analysis of the EF-hand domain-containing protein EFCAB14. The edited population is derived through CRISPR/Cas9-mediated gene disruption, creating a mixed pool of knockout genotypes that collectively abolish EFCAB14 expression. This format retains the biological heterogeneity of a non-clonal population, mitigating selection bias and providing a physiologically relevant background for investigating EFCAB14 function. Researchers can employ this system to interrogate calcium-dependent signaling cascades without the limitations of monoclonal isolates.
The 786-O cell line originates from a primary clear cell renal cell carcinoma (ccRCC) and serves as a well-established model of this malignancy. These cancer epithelial cells are VHL-negative, resulting in constitutive activation of hypoxia-inducible factor 1-alpha (HIF-1??) even under normoxia. The VHL deficiency mimics the genetic hallmark of the majority of sporadic ccRCCs, making 786-O an ideal host for dissecting molecular pathways relevant to renal tumorigenesis. Within this context, the interplay between HIF-1?? signaling and calcium-mediated pathways remains an active area of investigation, positioning the EFCAB14 knockout as a valuable tool for exploring calcium-dependent mechanisms in a disease-relevant setting.
EFCAB14 encodes an EF-hand domain-containing protein that putatively functions as an intracellular calcium sensor, transducing transient calcium signals into specific cellular outputs. In the context of the calcium signaling pathway, EFCAB14 likely responds to elevations in cytosolic calcium triggered by upstream regulators such as GPCR signaling or store-operated calcium entry through STIM1 and ORAI1 channels. As a calcium-binding protein, it may interact with and regulate downstream targets including calcium/calmodulin-dependent protein kinase kinases (CAMK kinases), calcineurin, and NFAT transcription factors. The protein is predicted to interact with calmodulin and other EF-hand proteins, integrating into a network that includes TRPC channels, CAMKII, and calcineurin. Disruption of EFCAB14 is hypothesized to broadly impair calcium-mediated signal transduction, potentially altering the balance of CAMK and calcineurin/NFAT cascades and thereby influencing cellular processes such as proliferation and survival.
In the 786-O VHL-negative ccRCC model, calcium signaling is frequently dysregulated and contributes to oncogenic properties including enhanced proliferation, migration, and resistance to apoptosis. The constitutive activation of HIF-1?? in these cells may intersect with calcium-dependent pathways, suggesting that EFCAB14 could play a role in mediating such cross-talk. By eliminating EFCAB14 in this polyclonal knockout population, researchers can directly assess how loss of this calcium sensor impacts HIF-1??-driven tumorigenic phenotypes. The polyclonal nature of the model preserves the genetic and phenotypic diversity of the original cell population, offering a robust platform for evaluating gene function while averaging out clone-specific artifacts. This system is particularly suited for studies aiming to link EFCAB14-dependent calcium signaling with ccRCC pathology.
This EFCAB14 knockout polyclonal cell product is designed for a wide range of functional assays critical to renal cancer research and calcium signaling studies. Typical applications include characterizing the role of EFCAB14 in calcium-dependent regulation of proliferation using MTT or colony formation assays, and assessing its impact on migration through transwell experiments. Calcium imaging can directly probe whether EFCAB14 loss alters intracellular calcium dynamics, while Western blotting and RT-qPCR enable confirmation of knockout and monitoring of downstream targets such as NFATc1 or phospho-CAMKII. Flow cytometry is suitable for quantifying apoptosis induction. Transcriptomic analyses via RNA-seq can reveal EFCAB14-dependent gene expression programs. These applications support drug target validation and mechanistic studies of calcium signaling in ccRCC. For additional details or custom inquiries, please contact Ascent Research.
