The HMGCL Knockout HT29 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout population with disrupted HMGCL gene in the HT29 colorectal adenocarcinoma cell line. This polyclonal format offers a heterogeneous editing pool without clonal bias, providing a robust loss-of-function model. The cells are generated via CRISPR/Cas9-mediated genome editing, capturing multiple editing events for robust phenotypic analysis.
The HT29 cell line, a human colorectal adenocarcinoma with epithelial morphology, is capable of enterocytic differentiation and harbors mutations in APC, TP53, and PIK3CA. These genetic alterations recapitulate key oncogenic pathways, making HT29 a relevant model for dissecting metabolic vulnerabilities in colorectal tumors. As an intestinal epithelial model for colorectal cancer research, HT29 cells are widely employed to study cancer metabolism and signaling in a genetically defined background.
HMGCL encodes a mitochondrial enzyme that cleaves HMG-CoA to acetyl-CoA and acetoacetate, essential for ketogenesis and leucine catabolism. HMGCL activity is crucial for generating ketone bodies during fasting and for processing leucine. The enzyme functions as a homodimer and interacts with HMGCS2 and AUH. Transcriptional activation by PPARA and regulation via glucagon and insulin through cAMP/PKA signaling control HMGCL expression. Downstream metabolites include acetoacetate, ??-hydroxybutyrate, and acetyl-CoA, which enter the TCA cycle or fatty acid synthesis. Pathway partners such as HMGCS2, BDH1, ACAT1, BCAT2, and AUH coordinate these metabolic processes.
In HT29 cells, HMGCL knockout eliminates ketone body production and leucine degradation, impairing metabolic adaptation to nutrient stress. Colorectal cancer cells often depend on these pathways during glucose limitation, so loss of HMGCL disrupts metabolic reprogramming critical for tumor cell survival. Disabling these processes forces reliance on alternative substrates, revealing potential therapeutic targets. This model is thus valuable for dissecting how cancer cells cope with metabolic challenges in the tumor microenvironment.
These polyclonal knockout cells are suited for studying metabolic reprogramming, ketone body metabolism, leucine catabolism, and nutrient stress responses. Assays include ketone body quantification, 13C-leucine metabolic flux analysis, cell viability under low glucose, Western blot and RT-qPCR for HMGCL, Seahorse mitochondrial stress tests, and apoptosis assays. These functional studies can be combined with genetic or pharmacological interventions to explore synthetic lethality or metabolic rescue mechanisms. The model aids in identifying metabolic vulnerabilities and evaluating cancer metabolic therapy targets. For further information, please contact technical support.
