The ELANE Knockout HT29 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population originating from the HT29 human colorectal adenocarcinoma line. This product disrupts the ELANE gene, which encodes neutrophil elastase, a serine protease central to extracellular matrix (ECM) degradation and inflammatory signaling. The polyclonal composition provides a diverse pool of edited cells, enabling functional studies without clonal selection bias. It serves as a loss-of-function model for examining ELANE-dependent mechanisms in human epithelial cells. Routine quality control validates genomic disruption at the population level.
HT29 cells exhibit an epithelial phenotype and are a standard model for intestinal barrier function and colorectal cancer biology. They form polarized monolayers and are frequently used to investigate tumor cell adhesion, migration, and drug permeability. Their epithelial origin is particularly relevant for dissecting interactions between cancer cells and the ECM. The availability of HT29-derived knockout tools, such as this ELANE knockout population, expands the capacity for targeted pathway analysis in colorectal carcinoma.
Neutrophil elastase, the ELANE gene product, is a chymotrypsin-like serine protease expressed in neutrophils and macrophages. Its activity is induced by inflammatory cytokines TNF-?? and IL-1??, and it is transcriptionally controlled by C/EBP and PU.1 downstream of G-CSF signaling. The protease directly cleaves ECM substrates including collagens, elastin, and proteoglycans, and contributes to tissue remodeling by activating matrix metalloproteinases (MMPs). Endogenous inhibitors SERPINA1 (alpha-1 antitrypsin), alpha-2 macroglobulin, and SLPI regulate its function. Additionally, elastase processes IL-8 and stimulates NF-??B, MAPK, and PI3K-Akt pathways, while integrin CD11b/CD18 links its activity to cell adhesion and migration.
Disruption of ELANE in HT29 cells abolishes neutrophil elastase activity, leading to attenuated ECM degradation and reduced activation of pro-invasive signaling. This is predicted to impair tumor cell migration and invasion, offering a model to study elastase-dependent colorectal cancer metastasis. The knockout also enables investigation of how loss of this protease reshapes the tumor microenvironment, including modulation of NF-??B- and STAT3-driven inflammatory circuits, and alters responsiveness to agents targeting ECM dynamics.
This knockout cell population is applicable to a variety of assays, including Transwell migration and invasion, immunoblotting for elastase, RT-qPCR for ELANE transcript levels, gelatin zymography for protease activity, and ELISA for cytokines such as IL-8. Research areas encompass colorectal cancer invasion and metastasis, ECM remodeling, inflammatory signaling, drug resistance, and therapeutic targeting of neutrophil elastase. Immunofluorescence detection of ECM components can complement functional analyses. For further information or to place an order, please contact Ascent Research.
