The EFHD1 Knockout 786-O Polyclonal Cells constitute a CRISPR/Cas9-edited polyclonal knockout cell population derived from the human 786-O renal cell carcinoma line, designed for targeted disruption of the EFHD1 gene. This polyclonal pool offers a heterogeneous loss-of-function model, avoiding artifacts associated with single-cell cloning and enabling robust population-level analyses of EFHD1-dependent biological processes.
The parental 786-O cell line is an established human clear cell renal cell adenocarcinoma model originating from a proximal tubule epithelial origin. These cells harbor a well-characterized mutation in the von Hippel-Lindau (VHL) tumor suppressor gene, resulting in constitutive stabilization of hypoxia-inducible transcription factors (HIF-1?? and HIF-2??) and persistent activation of HIF-target gene programs. This genetic background recapitulates hallmark features of renal cell carcinoma, including aberrant angiogenic signaling and metabolic reprogramming, making 786-O a highly relevant platform for studying kidney cancer biology.
EFHD1 encodes a cytosolic calcium-binding protein with two EF-hand motifs that function as calcium sensors. Upon calcium binding and stimulation by TNF-??, EFHD1 engages the mitochondrial apoptosis pathway by interacting with TRAF2 and Bcl-2 family members, promoting cytochrome c release and activation of caspase-9 and caspase-3. Simultaneously, it suppresses NF-??B signaling by impeding I??B degradation, thus limiting NF-??B nuclear translocation. EFHD1 also directly binds F-actin and regulates actin dynamics, influencing focal adhesion turnover and cell migration through modulation of FAK activity.
In VHL-mutant 786-O cells, constitutive HIF activation confers apoptosis resistance and a migratory phenotype. EFHD1 knockout further compromises calcium-dependent apoptotic signaling and releases NF-??B inhibition, likely enhancing survival, proliferation, and invasiveness. This polyclonal model thus allows dissection of EFHD1??s integrative role in calcium, HIF, and inflammatory pathways controlling renal cancer cell behavior.
These cells enable apoptosis assays (Annexin V/PI), Transwell migration/invasion, calcium imaging, and co-immunoprecipitation of EFHD1 complexes. They are also suitable for NF-??B reporter assays, phospho-kinase arrays, and RT-qPCR/Western blot validation. The polyclonal pool is ideal for unbiased drug resistance screening and protein interaction studies. For custom solutions, contact Ascent Research.
