The ACSS2 Knockout T-47D Polyclonal Cells are a CRISPR/Cas9-edited polyclonal population with targeted disruption of the ACSS2 gene in the T-47D human breast cancer epithelial cell line. This loss-of-function model provides a heterogeneous knockout background, avoiding clonal selection artifacts and enabling robust assessment of ACSS2 function across a diverse cell population.
T-47D cells, derived from a pleural effusion of infiltrating ductal carcinoma, are a widely used model of estrogen receptor-positive breast cancer. These hormone-responsive cells express estrogen and progesterone receptors, making them ideal for studying hormone-driven signaling and metabolic reprogramming. Their epithelial morphology and stable karyotype support a broad range of functional and biochemical analyses.
ACSS2 encodes acetyl-CoA synthetase 2, which converts acetate to acetyl-CoA, a key substrate for fatty acid synthesis and histone acetylation. The enzyme is transcriptionally regulated by SREBP1 and HIF-1??, and its activity is modulated by the PI3K/AKT/mTOR and AMPK pathways, as well as insulin/IGF-1 signaling. Acetyl-CoA generated by ACSS2 is utilized by downstream targets including fatty acid synthase (FASN) and histone acetyltransferases such as p300/CBP, linking nutrient availability to lipid biosynthesis and epigenetic modifications. Under hypoxic or nutrient-depleted conditions, ACSS2 sustains acetyl-CoA pools, promoting cancer cell proliferation and survival through lipid synthesis and chromatin remodeling.
In the T-47D breast cancer context, disruption of ACSS2 allows interrogation of acetate-dependent metabolic vulnerabilities. Estrogen receptor-positive breast cancers frequently depend on lipid metabolism and epigenetic regulation for growth and therapy resistance, processes potentially fueled by ACSS2-derived acetyl-CoA. The polyclonal knockout cells enable population-level studies of ACSS2 function, revealing how its loss impacts hormone-driven proliferation, metabolic flexibility, and the epigenetic landscape under microenvironmental stresses such as hypoxia or lipid scarcity.
These knockout cells are suited for a range of applications, including metabolic flux analysis with 14C/13C-acetate tracking, Western blotting for ACSS2 and histone H3 acetylation, RT-qPCR for ACSS2 and FASN expression, and Seahorse metabolic profiling. Further assays include lipid droplet staining, proliferation and migration/invasion assays, ChIP-seq for acetylated H3, acetate uptake measurements, and colony formation under lipid-depleted conditions. Such approaches support drug sensitivity profiling and assessment of metabolic vulnerabilities. For additional details, please contact Ascent Research.
