JOSD1 Knockout A-549 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal population derived from A-549 lung adenocarcinoma cells, providing a loss-of-function model for the JOSD1 deubiquitinase. The polyclonal knockout was generated by CRISPR/Cas9-mediated disruption of the JOSD1 gene, resulting in a heterogeneous pool of edited alleles without single-cell cloning. This format enables population-level studies where clonal variation is acceptable and simplifies experimental scalability. The cells are supplied as a viable stock for expansion under standard culture conditions.
The A-549 cell line originates from a 58-year-old male lung carcinoma patient and is a widely used model of alveolar basal epithelial adenocarcinoma. These adherent cells are extensively employed in cancer biology, drug screening, and toxicology due to their robust growth and disease-relevant characteristics. The tumor-derived background provides a clinically meaningful context for studying lung cancer mechanisms and therapeutic responses.
JOSD1 is a Josephin domain deubiquitinase that specifically cleaves Lys-63-linked polyubiquitin chains, modulating protein stability and signaling. It interacts with proteasome subunits, autophagy receptors such as p62/SQSTM1 and LC3, and other deubiquitinases. JOSD1 knockout leads to accumulation of K63-linked ubiquitin conjugates and protein aggregates, potentially dysregulating pathways including autophagy and the DNA damage response. Disruption of JOSD1 activity is therefore anticipated to alter protein turnover and stress responses.
In A-549 lung cancer cells, loss of JOSD1 deubiquitinase function may compromise proteostasis and influence cancer-associated signaling networks. The model can reveal roles of K63-ubiquitination in proliferation, survival under stress, and therapy resistance. It also serves as a tool to investigate mechanisms of protein aggregation and degradation, relevant to both oncogenesis and neurodegenerative disorders.
Applications include Western blotting for ubiquitin conjugates, deubiquitinase activity assays (ubiquitin-AMC), proteasome activity assays, immunofluorescence for aggregates, and co-immunoprecipitation of ubiquitinated substrates. These cells enable functional studies under proteotoxic stress, cell viability assays, and apoptosis analysis by flow cytometry. They are suited for target validation, biomarker discovery, and autophagy research. Contact Ascent Research for more information.
