This product is a polyclonal CRISPR/Cas9-edited knockout cell population derived from the 786-O human renal carcinoma cell line, with targeted disruption of the DNPEP gene. The knockout is achieved using CRISPR/Cas9-mediated genome editing to introduce functional gene disruption, creating a robust loss-of-function model. The polyclonal format ensures representation of diverse genetic edits, providing a population-level assessment of gene function and reducing artifacts associated with single-cell selection. This cell population is suitable for investigating the regulatory roles of aspartyl aminopeptidase in cancer biology.
The parental 786-O cell line is a well-characterized model of clear cell renal cell carcinoma (ccRCC), originally derived from a primary tumor. These cells exhibit constitutive activation of hypoxia-inducible pathways due to VHL inactivation, leading to stabilization of the transcription factor HIF1A and consequent upregulation of angiogenic factors such as VEGF. 786-O cells are widely employed to study ccRCC proliferation, apoptosis, and angiogenesis, making them an ideal host for probing aminopeptidase-mediated regulatory mechanisms.
DNPEP encodes aspartyl aminopeptidase, an enzyme that specifically cleaves N-terminal aspartate residues from peptide substrates, thereby modulating the bioactivity of peptide hormones and signaling molecules. In the 786-O background, DNPEP expression is regulated by transcription factors HIF1A and SP1, linking its function to hypoxic and growth-promoting pathways. Downstream, DNPEP-mediated processing alters levels of peptides such as angiotensin and bradykinin, impacting interactions with other aminopeptidases and influencing signaling networks. DNPEP activity intersects with the mTOR and AKT pathways, and it modulates BCL2 family-regulated apoptosis and VEGF-driven angiogenesis. Consequently, knockout of DNPEP disrupts this peptide processing hub, perturbing the balance between pro-survival and pro-apoptotic signals and attenuating angiogenic signaling.
In 786-O cells, loss of DNPEP function is expected to shift peptide hormone profiles, potentially reducing pro-angiogenic outputs or sensitizing cells to apoptotic stimuli. Given the centrality of VHL-HIF1A-VEGF signaling in ccRCC, DNPEP knockout may limit angiogenesis-dependent tumor growth and affect mTOR-mediated metabolic control. This model is significant for dissecting the contribution of aminopeptidase activity to ccRCC progression and for identifying substrate-dependent mechanisms that influence tumor cell proliferation and survival.
This polyclonal DNPEP knockout cell product supports a broad range of experimental applications. Researchers can confirm gene disruption by western blotting and RT-qPCR, and assess changes in downstream targets such as VEGF or BCL2 family members. Functional studies may employ MTT proliferation assays, migration and invasion assays, and tube formation assays to evaluate angiogenesis. Peptide cleavage assays allow direct measurement of enzymatic activity, while substrate identification studies elucidate processing pathways. The model is ideally suited for investigating DNPEP function in renal cancer, peptide hormone processing, and angiogenic signaling. For further technical details, please contact Ascent Research.
