The ITCH Knockout A-549 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout population of human A-549 cells with disruption of the ITCH gene. This loss-of-function model abolishes ITCH E3 ubiquitin ligase activity, enabling studies of substrate ubiquitination and degradation in a lung epithelial context. The polyclonal format provides a heterogeneous gene-edited pool that avoids clonal biases and is suitable for functional assays requiring population-level readouts. It serves as a vital resource for investigating ITCH-dependent regulation of signaling networks controlling apoptosis, proliferation, and immune function.
The A-549 cell line is a well-established model derived from human lung adenocarcinoma, exhibiting properties of alveolar basal epithelial cells. Widely used in respiratory research, these cells facilitate investigations of cancer biology, drug response, and epithelial function. They express key oncogenic mutations and display an epithelial phenotype, providing a relevant platform to study tumor cell signaling. Integrating ITCH knockout into this background enables focused analysis of E3 ligase-mediated control over pathways critical to lung cancer progression and maintenance.
ITCH encodes a HECT-type E3 ubiquitin ligase that transfers ubiquitin to target substrates, promoting their proteasomal degradation. Its activity is regulated by upstream factors including JNK phosphorylation and adaptor proteins NDFIP1 and N4BP1. Key substrates include p73, p63, c-FLIP, LATS1, and SMAD7, linking ITCH to the Hippo, TGF-??, Notch, and Wnt/??-catenin pathways. Through ubiquitination of these effectors, ITCH governs apoptosis, cell cycle progression, and inflammatory signaling. Dysregulation of ITCH has been implicated in autoimmune disorders, allergic diseases, and oncogenesis, making this knockout model a powerful system to dissect disease-relevant molecular mechanisms.
In A-549 cells, ITCH loss leads to stabilization of substrates like p73 and LATS1, promoting apoptosis and inhibiting proliferation. Concurrent accumulation of SMAD7 modulates TGF-?? signaling, while altered c-FLIP turnover sensitizes cells to death receptor activation. This polyclonal knockout population enables systematic investigation of how ITCH controls epithelial-mesenchymal transition, drug sensitivity, and immune evasion in lung adenocarcinoma. It also provides a tool to assess the interplay between ubiquitin-mediated degradation and NF-??B-driven inflammatory programs within the alveolar epithelial niche.
This product supports a wide range of biomedical research applications. Users can perform ubiquitination assays and western blotting to monitor substrate turnover, co-immunoprecipitation to probe ITCH interactors, and apoptosis assays with Annexin V/PI staining for cell death analysis. Flow cytometry and luciferase reporter assays enable quantitation of pathway activities. The cells are optimal for functional genomics screens, pharmacological studies, and mechanistic dissection of signaling networks in cancer biology, immunology, and respiratory disease. For inquiries, contact Ascent Research.
