The ELANE Knockout HCT 116 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal cell population derived from HCT 116 cells, engineered for disruption of the ELANE gene. This polyclonal format provides a heterogeneous pool of cells with varied loss-of-function mutations, reducing clonal bias and representing population-level effects.
HCT 116 is a well-characterized human male colorectal carcinoma cell line with epithelial adherent morphology, exhibiting microsatellite instability (MSI-H) and a KRAS G13D mutation, while retaining wild-type TP53. Widely utilized as a colorectal cancer model, HCT 116 cells facilitate the study of oncogenic signaling, drug response, and tumor microenvironment interactions. Their robust growth and genetic tractability make them an ideal chassis for CRISPR-mediated gene disruption.
ELANE encodes neutrophil elastase, a serine protease predominantly expressed in neutrophil granules but not in HCT 116 cells. Although absent in this line, neutrophil elastase is a key mediator of innate immunity, cleaving extracellular matrix proteins such as elastin, collagens, and fibronectin, and activating downstream effectors including MMP-9. Its expression is controlled by upstream regulators G-CSF, GM-CSF, IL-8, and TNF-??, along with transcription factors C/EBP?? and PU.1. ELANE activity is balanced by inhibitors like SERPINA1 (??1-antitrypsin), SLPI, and elafin, and it interacts with CD11b/CD18 integrin and proteoglycans. In neutrophil biology, ELANE contributes to NETosis and amplifies inflammatory cascades via NF-??B and TLR4 pathways, highlighting its multifaceted role in host defense and tissue remodeling.
Since HCT 116 cells do not endogenously express ELANE, the knockout background provides a clean platform to dissect neutrophil elastase-mediated effects when introduced exogenously, such as through co-culture with neutrophils or addition of recombinant enzyme. This model is particularly suited for evaluating the impact of elastase on tumor cell migration, invasion, and signaling, without confounding endogenous elastase activity. It also enables the study of protease-dependent modifications of the extracellular matrix and integrin signaling in a controlled setting, facilitating the identification of specific substrates and downstream pathways.
Researchers can employ these ELANE Knockout HCT 116 Polyclonal Cells to investigate the role of neutrophil elastase in the tumor microenvironment, screen for small-molecule elastase inhibitors, and analyze protease-driven ECM remodeling using transwell migration/invasion assays and cytokine profiling. Co-culture systems with primary neutrophils or differentiated neutrophil-like cells allow assessment of tumor-immune cell crosstalk, NETosis induction, and inflammatory mediator release. The cells also serve as an in vitro model for conditions such as cyclic neutropenia or acute lung injury where aberrant elastase activity is implicated. Standard validation techniques include western blotting and RT-qPCR to confirm gene disruption, while functional assays like elastase activity assays and proliferation analysis provide insight into downstream consequences. For further information, contact Ascent Research.
