The GNPAT Knockout A-549 Polyclonal Cells constitute a CRISPR/Cas9-edited polyclonal knockout population derived from the human A-549 lung adenocarcinoma cell line. The targeted disruption of GNPAT generates a heterogeneous pool of cells collectively lacking functional GNPAT expression. This population model is designed for investigating ether lipid biosynthesis, peroxisomal metabolism, and their implications in cancer and other diseases.
The A-549 cell line originated from carcinomatous lung tissue of a 58-year-old Caucasian male and serves as a standard in vitro model for type II pneumocytes and lung adenocarcinoma. These cells exhibit intact peroxisomal machinery and are amenable to assays probing metabolic activity, invasion, and migration capabilities. GNPAT knockout in this setting enables context-specific studies of ether phospholipid metabolism and its disruption in lung cancer.
GNPAT (glyceronephosphate O-acyltransferase) is a peroxisomal enzyme that initiates ether phospholipid synthesis by acylating dihydroxyacetone phosphate (DHAP) to form 1-acyl-DHAP. This first step is critical for the generation of plasmalogens, which are abundant in cell membranes and act as antioxidants. GNPAT transcription is regulated by PPAR??, a central lipid metabolism mediator, and SREBP1, a master regulator of fatty acid and sterol synthesis. Downstream, 1-acyl-DHAP is processed by AGPS and other peroxisomal matrix enzymes, requiring PEX7 for enzyme import and functioning in coordination with FAR1.
Loss of GNPAT function in A-549 cells allows the dissection of ether phospholipid-dependent processes in a lung adenocarcinoma background. This knockout model is instrumental for mimicking rhizomelic chondrodysplasia punctata type 2 (RCDP2), a disorder linked to GNPAT mutations and plasmalogen deficiency, and for exploring how compromised ether lipid synthesis impacts cancer cell phenotypes such as proliferation, metabolic flexibility, and invasive capacity.
Typical experimental workflows with these cells include lipidomic analysis to profile ether phospholipid changes, Western blotting and RT-qPCR for expression verification, resazurin-based viability tests, immunofluorescence staining for peroxisomal or cytoskeletal markers, and migration/invasion assays to evaluate metastatic behavior. The knockout population also facilitates drug screening targeting ether lipid pathways and mechanism-of-action studies. For additional technical specifications or inquiries, please contact Ascent Research.
