HSDL2 Knockout HT29 Polyclonal Cells are a CRISPR/Cas9-edited population of HT29 human colorectal adenocarcinoma cells with disruption of the HSDL2 gene, providing a loss-of-function model for studying mitochondrial lipid metabolism and signaling. The polyclonal format preserves editing diversity, enabling functional genomics without clonal selection bias, while retaining the parental HT29 epithelial morphology and genetic background for direct comparison with wild-type controls.
The HT29 host cell line was derived from a human colon adenocarcinoma and exhibits epithelial adherent growth with mutations in APC and TP53, coupled to dysregulated Wnt/??-catenin and RAS pathways. Widely used for intestinal cancer research, HT29 cells can undergo differentiation, offering a tractable system to examine heterogeneous tumor cell behaviors and metabolic adaptations in colorectal cancer.
HSDL2 encodes a mitochondrial short-chain dehydrogenase/reductase that catalyzes the conversion of phosphatidic acid to diacylglycerol, thereby regulating phospholipid homeostasis and the generation of signaling-competent lipids. Its expression is transcriptionally controlled by the SP1 transcription factor and is likely modulated by nuclear receptor pathways. Downstream, HSDL2 activity impinges on the PI3K-AKT and MAPK/ERK cascades: loss of HSDL2 reduces AKT and ERK1/2 phosphorylation, leading to decreased cyclin D1 levels and attenuated Rho GTPase signaling, including RhoA. Interacting partners include mitochondrial membrane proteins and enzymes involved in phosphatidate metabolism, positioning HSDL2 at a critical node linking lipid metabolic flux to proliferative and migratory signaling.
In the HT29 colorectal adenocarcinoma model, HSDL2 knockout produces a marked reduction in proliferation and migration, consistent with its role in driving oncogenic lipid signaling. The polyclonal cells display diminished AKT and ERK1/2 activation and reduced cyclin D1 expression, resulting in delayed cell cycle progression. These deficits highlight HSDL2 as a potential metabolic vulnerability in colorectal cancer and provide a comparative context for studying related mechanisms in breast cancer and metabolic disorders.
Applications include western blotting for phospho-AKT and phospho-ERK1/2 to assess signaling, MTT/BrdU proliferation assays, transwell migration/invasion studies, and lipidomic profiling of phosphatidic acid/diacylglycerol levels. RT-qPCR and flow cytometry can quantify downstream targets and cell cycle distribution. The polyclonal knockout population is well-suited for drug target validation and high-content genetic or chemical screens. For additional information, please contact Ascent Research.
