The ELANE Knockout HAP1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population for the ELANE gene in the HAP1 human cell line. This polyclonal population, generated by CRISPR/Cas9-mediated gene disruption, contains a diverse array of loss-of-function mutations at the ELANE locus, avoiding clonal artifacts. The model provides an ideal system for studying neutrophil elastase biology in a simplified genetic environment.
The HAP1 cell line is a near-haploid human cell line derived from the KBM-7 chronic myeloid leukemia line, exhibiting fibroblastoid morphology. Its haploid state simplifies gene targeting, as a single allele disruption yields a functional knockout. HAP1 is widely adopted for CRISPR-based functional genomics, offering robust growth and compatibility with various assays.
ELANE encodes neutrophil elastase, a serine protease stored in azurophil granules that degrades extracellular matrix components such as elastin, collagen IV, and fibronectin, as well as processes cytokines IL-1?? and TNF. It is transcriptionally regulated by C/EBP?? and PU.1, and is responsive to G-CSF, with Runx1 also contributing to its expression. Endogenous inhibitors include SERPINA1 (??1-antitrypsin), SLPI, and A2M. Functionally, neutrophil elastase works in concert with cathepsin G, proteinase 3, and azurocidin within azurophil granules, driving pathogen clearance and neutrophil extracellular trap formation. ELANE knockout thus compromises innate immune effector functions and tissue remodeling.
In the HAP1 background, ELANE knockout enables dissection of protease-dependent pathways without neutrophil-specific factors. This model is relevant for investigating the molecular basis of severe congenital neutropenia and cyclic neutropenia, frequently associated with ELANE mutations, and for emphysema studies where elastase-inhibitor imbalance is key.
This polyclonal knockout cell population is suited for elastase activity assays, Western blotting of downstream targets like E-cadherin, degranulation assays, and flow cytometry. Researchers can use it for cell migration/invasion assays to study protease-mediated motility, drug screening for neutropenia, and testing protease inhibitor efficacy. For technical inquiries, contact Ascent Research.
