The HSDL1 Knockout HT29 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population designed for functional studies of the HSDL1 gene in a human colorectal adenocarcinoma background. This loss-of-function model enables investigation of HSDL1-dependent cellular processes through target-gene disruption in a heterogeneous pool of edited cells, reflecting population-level gene ablation effects. The polyclonal format provides a robust system for studying general HSDL1 knockout phenotypes within the context of colorectal cancer epithelial biology, suitable for both mechanistic and applied research applications.
The HT29 host cell line is a well-characterized model derived from a human colorectal adenocarcinoma with epithelial differentiation. It harbors oncogenic mutations in APC, TP53, and KRAS, while retaining mismatch repair proficiency and an invasive phenotype. HT29 cells form polarized monolayers expressing features of intestinal epithelial cells, making them a relevant system for studying intestinal biology, oncogenic signaling, and tumor cell metabolism. Their genetic profile recapitulates key aspects of colorectal cancer progression, providing a clinically pertinent setting for targeted gene disruption.
HSDL1 encodes a short-chain dehydrogenase/reductase enzyme that acts as a putative oxidoreductase involved in steroid and lipid metabolism. The protein plays a key role in regulating fatty acid oxidation and lipid droplet dynamics, functions that are integrated within the PPAR signaling network. HSDL1 is transcriptionally regulated by PPARG, SREBF1, NR1H3, and insulin, while it influences downstream targets such as ACADM, CPT1A, PLIN2, PPARA, and FABP4. Through interactions with cofactors NAD+ and NADP+ and lipid droplet-associated proteins PLIN2 and FABP4, HSDL1 contributes to the coordination of lipid storage and energy metabolism pathways.
In HT29 colorectal cancer cells, HSDL1 knockout is expected to disrupt lipid droplet formation and impair fatty acid oxidation, thereby altering PPAR signaling and metabolic homeostasis. This disruption may diminish the proliferative and migratory capacity of these tumor cells, linking HSDL1 loss to reduced tumorigenic properties. The model is particularly valuable for dissecting the intersection of lipid metabolism and oncogenic signaling in colorectal cancer, including the investigation of obesity-related and metabolic syndrome-associated disease mechanisms, as well as cancer cachexia.
Research applications include detailed metabolic phenotyping using Oil Red O and BODIPY staining to visualize neutral lipids and lipid droplets, fatty acid oxidation assays, and Seahorse metabolic flux analysis. Complementary approaches such as western blotting for PPARG and PLIN2, RT-qPCR for HSDL1 and ACADM, cell proliferation, migration, and colony formation assays facilitate comprehensive functional studies. This polyclonal knockout cell population is suitable for drug sensitivity screening and metabolic reprogramming research in colorectal cancer models. For additional technical specifications or ordering information, please contact Ascent Research.
