The CBR4 Knockout 786-O Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the 786-O human renal cell adenocarcinoma line, designed for disruption of the CBR4 gene. This polyclonal pool provides a versatile loss-of-function model to study CBR4-dependent metabolic processes without clonal selection.
The parental 786-O cell line originates from a primary clear cell renal cell carcinoma (ccRCC) in a 58-year-old male and harbors a mutant VHL tumor suppressor. VHL deficiency leads to constitutive HIF activation, altering cellular metabolism and establishing 786-O as a key model for ccRCC research.
CBR4 encodes a short-chain dehydrogenase/reductase that catalyzes the NADPH-dependent conversion of 3-ketoacyl-ACP to 3-hydroxyacyl-ACP in mitochondrial fatty acid synthesis (mtFAS). This step is critical for lipoic acid biosynthesis and the function of mitochondrial 2-oxoacid dehydrogenases. CBR4 functions downstream of malonyl-CoA-ACP transacylase (MCAT) and 3-oxoacyl-ACP synthase (OXSM), and upstream of enoyl-ACP reductase (MECR), with transcriptional regulation by SREBP1 and PPARGC1A. Loss of CBR4 disrupts lipoylation of proteins like DLAT and DLST, impairing TCA cycle activity and respiratory chain function.
In the VHL-mutant 786-O context, CBR4 knockout is expected to perturb mitochondrial energy homeostasis and lipid metabolism, potentially uncovering metabolic vulnerabilities specific to ccRCC. This model allows investigation of mtFAS contributions to tumor cell metabolism and interactions with HIF-driven pathways.
Applications include functional dissection of CBR4 in ccRCC metabolism, target validation for metabolic therapies, and metabolic profiling via Seahorse analysis, lipidomics, and RNA-seq. Researchers can assess lipoylation by Western blot, quantify mtFAS gene expression by RT-qPCR, and evaluate proliferation. For inquiries, contact Ascent Research.
