The GNPAT Knouckout Jurkat Polyclonal Cells consist of a CRISPR/Cas9-edited polyclonal knockout cell population originating from the Jurkat human T lymphocyte line. This product delivers targeted disruption of the GNPAT gene, which encodes the peroxisomal enzyme glyceronephosphate O-acyltransferase, across a heterogeneous pool of cells. The polyclonal format preserves genetic diversity among loss-of-function alleles, reducing clonal artifacts and providing a more physiologically relevant model for studying ether lipid metabolism. Cells are supplied cryopreserved and can be rapidly expanded for downstream applications.
Jurkat cells are an immortalized human T lymphocyte line derived from the peripheral blood of a patient with acute T cell leukemia. They are widely employed as a model for T cell receptor (TCR) signaling, T cell activation, and leukemogenesis due to their well-characterized response to immunological stimuli such as anti-CD3 and phorbol esters. Their robust growth, ease of culture, and amenability to genetic modification make them a preferred system for CRISPR/Cas9-mediated knockout studies in an immune-relevant cellular background.
GNPAT functions as the gatekeeper of ether lipid biosynthesis, catalyzing the peroxisomal acylation of dihydroxyacetone phosphate (DHAP) to 1-acyl-DHAP. This step is essential for the subsequent action of alkylglycerone phosphate synthase (AGPS) and the production of plasmalogens, along with the signaling molecule platelet-activating factor (PAF). GNPAT activity is modulated by PPAR??-mediated transcriptional control and is dependent on the PEX7 import receptor for peroxisomal targeting. Within the organelle, it partners with AGPS and utilizes acyl-CoA substrates, positioning GNPAT at the intersection of peroxisomal metabolism, membrane lipid biogenesis, and intercellular lipid signaling.
In Jurkat T lymphocytes, disruption of GNPAT abolishes plasmalogen synthesis, leading to altered plasma membrane composition that may influence lipid raft integrity and TCR signal transduction. This knockout model thus enables detailed examination of how ether lipid deficiency impacts T cell activation thresholds, calcium signaling, and cytokine production. Additionally, because inactivating GNPAT mutations cause rhizomelic chondrodysplasia punctata type 2 (RCDP2), a severe peroxisomal disorder, these cells serve as a human lymphocyte-based platform for exploring RCDP2 pathology and evaluating potential therapeutic compounds.
The polyclonal knockout population is suited for a variety of experimental approaches, including western blot and RT-qPCR for gene expression validation, mass spectrometry?Cbased lipidomics to quantify plasmalogen and PAF levels, and peroxisomal enzyme activity assays. Functional analyses can incorporate flow cytometric assessment of T cell activation markers, TCR stimulation assays, and metabolic labeling of ether lipids. Drug screening for RCDP2 and studies of metabolic-immune crosstalk are additional applications. For technical inquiries or to request a quote, please contact Ascent Research.
