The KDSR Knockout A-549 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal cell population in which the KDSR gene encoding 3-ketodihydrosphingosine reductase has been disrupted, creating a loss-of-function model for sphingolipid biosynthesis studies. The polyclonal format represents a heterogeneous pool of knockout alleles, providing a population-level assessment of gene function relevant to cancer research.
The host A-549 cell line originates from human lung adenocarcinoma and serves as an in vitro model for alveolar type II epithelium. These adherent epithelial cells are widely employed in lung cancer and respiratory biology studies, retaining oncogenic features such as dysregulated proliferation and apoptosis resistance.
KDSR catalyzes the NADPH-dependent reduction of 3-ketodihydrosphingosine to dihydrosphingosine, a committed step in de novo sphingolipid synthesis. Dihydrosphingosine is subsequently converted to ceramide by ceramide synthases (CERS1-6), and ceramide is further metabolized to sphingomyelin and sphingosine-1-phosphate (S1P). The pathway is regulated by transcription factors SREBP1 and PPAR?? and responds to nutrient status. Knockout of KDSR disrupts production of essential sphingolipid intermediates, potentially altering membrane architecture and signaling through ceramide as a pro-apoptotic mediator and S1P as a survival factor.
In A-549 lung adenocarcinoma cells, sphingolipid metabolism supports tumor growth and stress adaptation. Disabling KDSR in these cells is predicted to reduce ceramide and S1P levels, impacting apoptosis sensitivity, cell viability, and migratory capacity. This polyclonal knockout system enables robust assessment of sphingolipid pathway dependency without clonal selection bias, mimicking heterogeneous tumor environments.
Applications include sphingolipid profiling by LC-MS/MS, ceramide quantification, S1P ELISA, cell viability and apoptosis assays (MTT, Annexin V/PI), migration assays, and metabolic flux analysis with labeled palmitate. Downstream signaling can be analyzed by western blot for cleaved caspase-3 or transcriptomics. This model is suitable for studying sphingolipid roles in lung adenocarcinoma progression, drug resistance, and metabolic reprogramming. Contact Ascent Research for ordering and technical information.
