ACSS2 Knockout HT29 Polyclonal Cells provide a CRISPR/Cas9-edited polyclonal knockout cell population designed for loss-of-function studies of ACSS2 in a human colorectal adenocarcinoma background. The product is generated through CRISPR/Cas9-mediated gene disruption in the HT29 cell line, yielding a heterogeneous pool of edited cells without single-cell cloning. This polyclonal format preserves population-level heterogeneity while ensuring robust target-gene ablation, enabling functional interrogation of ACSS2-dependent processes without clonal selection artifacts. The knockout cells are suitable for a broad range of in vitro assays, including metabolic flux analysis, epigenetic profiling, and proliferation studies, and serve as a critical tool for dissecting the role of acetate metabolism in cancer biology.
HT29 cells, derived from a human colorectal adenocarcinoma, represent a widely used epithelial cancer model characterized by oncogenic mutations in APC, TP53, and KRAS, and retain a partially differentiated phenotype with the capacity for mucin production. These cells are extensively employed in studies of colorectal cancer progression, metabolic adaptation, and chemoresistance. Their well-documented growth requirements and metabolic dependencies make them an ideal host for investigating ACSS2 function, particularly in the context of nutrient stress and lipid biosynthesis. The HT29 background enables researchers to elucidate how ACSS2-mediated acetate utilization contributes to the malignant phenotype of colorectal cancer.
ACSS2 encodes acetyl-CoA synthetase short-chain family member 2, a ligase that catalyzes the ATP-dependent conversion of acetate to acetyl-CoA, a central metabolite at the intersection of energy metabolism, lipid synthesis, and protein acetylation. Transcriptionally activated by SREBP1, SREBP2, ChREBP, and HIF1A in response to insulin, acetate availability, and hypoxic conditions, ACSS2 is a key node in the acetate metabolic pathway. Its product, acetyl-CoA, serves as substrate for ACC, FASN, and HMGCR in de novo lipogenesis, and for histone acetyltransferases, thereby directly influencing gene expression through histone H3 acetylation. ACSS2 also interacts with metabolic regulators such as AMPK and ACLY, and is functionally intertwined with the TCA cycle, underscoring its role in maintaining metabolic homeostasis and epigenetic landscapes under fluctuating nutrient conditions.
In HT29 cells, ACSS2 supports survival and proliferation under metabolic stress by providing acetyl-CoA for lipid synthesis and histone modification, processes essential for sustaining tumor growth. The knockout model disrupts this metabolic conduit, enabling investigation of how colorectal cancer cells rewire their metabolism in the absence of exogenous acetate utilization. This is particularly relevant given the elevated expression of ACSS2 in many cancers and its correlation with poor prognosis. By eliminating ACSS2 activity, researchers can assess compensatory pathways, alterations in lipid profiles, changes in global histone acetylation, and effects on cell cycle progression, providing insights into the therapeutic potential of targeting acetate metabolism in colorectal adenocarcinoma.
This product is optimally suited for a range of applications in cancer metabolism and epigenetic research. Typical experimental workflows include Seahorse metabolic flux assays to measure oxygen consumption and extracellular acidification, metabolomics and lipidomics to profile acetyl-CoA pools and lipid species, and histone acetylation assays via Western blotting or ChIP-qPCR to evaluate epigenetic changes. Cell proliferation and colony formation assays can reveal functional consequences of ACSS2 loss, while RNA-seq enables transcriptomic analysis. The polyclonal knockout cells are also valuable for drug target validation and screening of small-molecule inhibitors targeting ACSS2 or downstream pathways. For further technical specifications, please contact Ascent Research.
