The ELANE Knockout 786-O Polyclonal Cells constitute a CRISPR/Cas9-edited polyclonal knockout population from the 786-O human renal cell adenocarcinoma line. This product delivers a heterogeneous loss-of-function model wherein ELANE disruption was achieved through CRISPR/Cas9-mediated gene editing, generating a mixed pool of cells with varied mutations. As a polyclonal sample, it represents a practical alternative to single-cell-derived clones, enabling functional genomics studies without clonal selection. The knockout is optimized for exploring non-hematopoietic roles of neutrophil elastase, particularly within the renal cancer microenvironment.
The parental 786-O cell line was established from a primary clear cell renal cell carcinoma (ccRCC) and carries a VHL tumor suppressor mutation, characteristic of the majority of ccRCC cases. This line is extensively used to model ccRCC biology, including hypoxia signaling dysregulation, metabolic reprogramming, and metastatic progression. Its adherent epithelial phenotype and stable culturing properties make it a robust platform for genetic manipulation and downstream functional analyses.
ELANE encodes neutrophil elastase, a serine protease classically involved in innate immunity through ECM protein degradation and inflammatory mediator processing. Beyond neutrophils, ELANE exhibits non-canonical expression in certain cancers, potentially modulating tumor progression. It functions downstream of regulators such as G-CSF (CSF3), TNF-alpha, and IL-8, and is inhibited by SERPINA1 (??1-antitrypsin) and SLPI. Its substrates include elastin, collagen, fibronectin, and pro-IL-1??, and it interacts with cathepsin G, myeloperoxidase, and protease-activated receptors. ELANE knockout accordingly disrupts ECM turnover, cytokine maturation (e.g., IL-1?? activation), and related signaling cascades, impacting processes like neutrophil extracellular trap formation and serine protease pathways. These perturbations can modify cellular behaviors tied to inflammation and tissue remodeling.
In the 786-O renal carcinoma context, ELANE knockout enables dissection of its non-canonical functions within the tumor microenvironment. Given the VHL-mutant background, this model allows interrogation of crosstalk between ELANE-mediated proteolysis and hypoxia-driven pathways. Loss of ELANE may affect ECM degradation and cytokine processing, potentially influencing tumor cell invasion, migration, and stromal interactions. This tool thus facilitates studies on how cancer-cell-intrinsic ELANE activity contributes to metastatic potential and drug response, opening avenues for therapeutic targeting in ccRCC.
This polyclonal knockout product supports a variety of experimental workflows. Validation of ELANE ablation can be performed by western blotting and RT-qPCR, while functional consequences are assessable via gelatin zymography for protease activity and ELISA for cytokine secretion changes. Phenotypic assays include Matrigel invasion and migration tests, cell proliferation measurements, and drug sensitivity profiling with chemotherapeutics. Transcriptome-wide analysis via RNA-seq can identify pathway-level alterations. Additionally, the cells are suitable for screening neutrophil elastase inhibitors and for modeling congenital neutropenia-associated pathways in a non-hematopoietic setting. For further technical details or to place an order, please contact Ascent Research.
