The DNAJC15 Knockout 786-O Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the human 786-O clear cell renal cell carcinoma line. This product consists of a mixed population of cells carrying targeted disruptions in the DNAJC15 gene, generated via CRISPR/Cas9-mediated gene disruption. The polyclonal knockout format avoids clonal artifacts and allows the study of gene function in a genetically diverse cellular background, making it suitable for broad phenotyping experiments. By ablating DNAJC15 expression across the population, these cells serve as a powerful tool for dissecting the mitochondrial and apoptotic pathways influenced by this co-chaperone.
The 786-O cell line is a well-characterized model of clear cell renal cell carcinoma (ccRCC) with biallelic VHL inactivation, resulting in constitutive HIF-1?? stabilization. This VHL deficiency drives a pseudohypoxic transcriptional program that promotes angiogenesis and metabolic reprogramming. 786-O cells are widely used to investigate ccRCC tumorigenesis, metastasis, and drug responses. Their VHL-mutant background creates a unique context for examining how mitochondrial co-chaperones like DNAJC15 impact cancer cell fitness under conditions of chronic HIF activation.
DNAJC15 is a mitochondrial inner membrane co-chaperone that partners with HSPA9 (mortalin) and the TIM23 translocase component TIMM17A to facilitate the import and folding of nuclear-encoded preproteins. It is specifically required for respiratory complex I assembly by mediating the import of subunits such as NDUFS3. Additionally, DNAJC15 interacts with Bcl-2 family proteins, including Bax, to regulate mitochondrial outer membrane permeabilization and apoptosis. Its expression is controlled by HSF1 and mitochondrial stress signals, and epigenetic silencing via DNA methylation occurs in some cancers. In the 786-O context, DNAJC15 knockout disrupts mitochondrial proteostasis, impairs respiratory function, and sensitizes cells to apoptosis.
In VHL-null 786-O cells, constitutive HIF activation suppresses mitochondrial respiration and promotes glycolysis, creating a metabolic vulnerability. Knockout of DNAJC15 exacerbates mitochondrial dysfunction by impairing protein import and complex I assembly, potentially compromising tumor cell survival. The resulting increased apoptosis sensitivity, particularly to BH3 mimetics, highlights the model’s utility for uncovering synthetic lethal interactions. This polyclonal knockout population thus offers a physiologically relevant tool to dissect the role of mitochondrial co-chaperones in ccRCC fitness.
These cells enable functional studies of mitochondrial import dynamics, respiratory chain complex assembly, and metabolic flux using Seahorse analysis. Apoptosis signaling can be monitored by flow cytometry (Annexin V), and mitochondrial morphology assessed by immunofluorescence. Co-immunoprecipitation assays allow profiling of DNAJC15 interactors, while drug sensitivity screens can evaluate agents targeting mitochondrial function or Bcl-2 family proteins. The knockout model is well-suited for investigating mitochondrial stress responses in ccRCC. For further details, contact Ascent Research.
