The LPCAT2 Knockout HT-29 Cell Line is a CRISPR/Cas9-edited knockout cell line derived from HT-29 human colorectal adenocarcinoma cells. This product features targeted disruption of the LPCAT2 gene, encoding lysophosphatidylcholine acyltransferase 2, providing a defined loss-of-function model for studying phospholipid metabolism and inflammatory signaling. The knockout eliminates functional LPCAT2 protein, enabling precise investigation of its role in membrane remodeling and eicosanoid biosynthesis without off-target effects associated with pharmacological inhibitors.
The HT-29 parental cell line is a widely used human epithelial colorectal adenocarcinoma model with epithelial morphology. It retains key characteristics of intestinal epithelium, including polarized monolayer formation and expression of differentiation markers, and is responsive to inflammatory stimuli such as TNF-?? and LPS. These features make HT-29 an appropriate host for examining the role of phospholipid metabolism in intestinal inflammation and colorectal cancer biology. The LPCAT2 knockout in this background allows for focused studies on how LPCAT2-dependent lipid pathways influence tumor cell behavior.
LPCAT2 catalyzes the acylation of lysophosphatidylcholine with arachidonoyl-CoA to produce arachidonoyl-phosphatidylcholine, a critical step in the Lands’ cycle. This reaction supplies substrates for phospholipase A2-mediated release of arachidonic acid, which is then converted by cyclooxygenase and lipoxygenase into pro-inflammatory eicosanoids such as prostaglandins and leukotrienes. LPCAT2 expression is activated by inflammatory signals like LPS and TNF-?? via NF-??B and AP-1 transcription factors. It also contributes to platelet-activating factor biosynthesis. Thus, LPCAT2 links membrane phospholipid remodeling to lipid mediator production, positioning it at the intersection of metabolism and inflammation.
Disruption of LPCAT2 in HT-29 cells is anticipated to reduce arachidonoyl-phosphatidylcholine levels, limiting free arachidonic acid availability and dampening eicosanoid production. This alteration can affect membrane composition, signaling microdomains, and downstream cellular responses. Given the role of lipid mediators in colorectal tumor progression and inflammation, this knockout model enables dissection of LPCAT2-dependent contributions to processes such as cell proliferation, migration, and drug resistance. It also offers a tool to examine how altered phospholipid profiles impact the tumor microenvironment and response to inflammatory cytokines.
Key applications include lipidomics mass spectrometry to profile phospholipid changes, RT-qPCR and western blotting for gene and protein expression analysis, and functional assays like cell proliferation, migration, and invasion. Eicosanoid profiling by ELISA and phospholipase A2 activity assays can quantify inflammatory mediator output. This cell line is valuable for research into lipid metabolism in colorectal cancer, inflammatory signaling, and drug resistance. For more information, please contact Ascent Research.
