The HLCS Knockout HCT 116 Polyclonal Cells product provides a CRISPR/Cas9-edited polyclonal cell population derived from the HCT 116 human colorectal carcinoma cell line, featuring targeted disruption of the HLCS gene. This gene-edited pool offers a genetically heterogeneous loss-of-function model for holocarboxylase synthetase (HLCS) research, avoiding the need for clonal isolation and enabling immediate use in population-level functional studies.
The HCT 116 parental cell line is a well-characterized human colorectal carcinoma model that harbors an activating KRAS G13D mutation and is microsatellite stable (MSS), making it particularly relevant for studying oncogene-driven metabolic reprogramming. These adherent epithelial cells are routinely cultured and are amenable to a wide range of genetic manipulation and biochemical assays. Their robust growth characteristics and defined genetic background provide a consistent platform for dissecting signaling and metabolic pathways relevant to colorectal cancer pathogenesis.
HLCS encodes holocarboxylase synthetase, which catalyzes the ATP-dependent biotinylation of apocarboxylases, thereby activating key metabolic enzymes. Its critical substrates include acetyl-CoA carboxylase (ACC), pyruvate carboxylase (PC), propionyl-CoA carboxylase (PCC), and 3-methylcrotonyl-CoA carboxylase (MCC). These biotin-dependent carboxylases are essential for fatty acid synthesis (ACC), gluconeogenesis and anaplerosis (PC), and the degradation of odd-chain fatty acids and leucine (PCC and MCC). HLCS activity is regulated by biotin availability, which is controlled by the sodium-dependent multivitamin transporter (SMVT), establishing a direct link between cellular nutrient uptake and metabolic enzyme activation. Disruption of HLCS therefore eliminates the biotinylation and activity of these carboxylases, leading to broad metabolic consequences.
In the HCT 116 colorectal carcinoma background, loss of HLCS function intersects with the oncogenic KRAS G13D mutation, which itself reprograms cellular metabolism. The combined effects may deplete lipogenic acetyl-CoA carboxylase activity and impair anaplerotic pyruvate carboxylase flux, potentially sensitizing cells to metabolic stress. This polyclonal knockout model is thus ideal for studying the functional interplay between constitutive KRAS signaling and the biotin-dependent metabolic network, and for identifying synthetic lethal interactions in colorectal cancer metabolism.
Key applications include streptavidin-based western blotting for global protein biotinylation, specific carboxylase activity assays, and metabolic profiling using LC-MS or Seahorse analysis to quantify fatty acid synthesis and oxidative metabolism. Proliferation assays under varying biotin concentrations can dissect nutrient sensing pathways. This tool supports gene function validation and cancer metabolism studies in a genetically defined colorectal carcinoma model. For further details or assistance, please contact Ascent Research.
