The ELANE Knockout A-549 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal human cell population designed for disruption of the ELANE gene in the A-549 lung adenocarcinoma epithelial line. This polyclonal knockout model introduces loss-of-function mutations through CRISPR/Cas9-mediated gene editing, yielding a heterogeneous population of cells with ablated neutrophil elastase expression. The product enables researchers to investigate the non-neutrophil functions of ELANE in an epithelial context, offering a versatile tool for dissecting protease-dependent signaling and matrix remodeling pathways relevant to lung biology and disease.
The A-549 cell line was established from lung adenocarcinoma tissue of a 58-year-old Caucasian male. These cells display characteristics of human alveolar type II epithelium and serve as a widely used model for lung adenocarcinoma pathogenesis, epithelial biology, and respiratory drug testing. A-549 cells form confluent epithelial monolayers, express surfactant-associated proteins, and respond to inflammatory stimuli, making them suitable for studying how protease loss affects epithelial behavior and tumor microenvironment interactions.
ELANE encodes neutrophil elastase, a serine protease primarily expressed in neutrophil granules with roles in innate immunity. The enzyme degrades bacterial proteins and remodels the extracellular matrix (ECM) by cleaving elastin and collagens. It also processes cytokines IL-8 and TNF-?? and modulates immune adhesion via ICAM-1 and CD14. Endogenous inhibitors ??1-antitrypsin (SERPINA1) and secretory leukoprotease inhibitor (SLPI) regulate its activity. ELANE transcription is driven by CSF3, C/EBP??, and PU.1. Dysregulated elastase activity contributes to tissue damage in COPD, emphysema, and ARDS.
Eliminating ELANE in A-549 epithelial cells allows exploration of the protease’s functions outside neutrophils. It permits dissection of epithelial-derived matrix remodeling, cytokine processing, and the interplay between ELANE and its inhibitors in a lung carcinoma background. Researchers can assess how loss of neutrophil elastase alters ECM degradation dynamics, cytokine profiles, and cell migration, distinguishing neutrophil-dependent versus epithelial-intrinsic contributions in tumor progression and inflammation.
These polyclonal KO cells are suited for western blotting and RT-qPCR to confirm gene disruption, elastin and collagen degradation assays, and ELISA-based cytokine quantification (IL-8, TNF-??). Functional assays include transwell migration/invasion, proliferation analysis, and RNA-seq for transcriptome profiling. The model supports drug screening for elastase inhibitors, investigation of lung cancer-ECM interactions, and mechanistic studies of respiratory diseases. For further information, contact Ascent Research.
