The ACOT13 Knockout HT29 Polyclonal Cells constitute a CRISPR/Cas9-edited polyclonal knockout cell population engineered to disrupt the ACOT13 gene in the HT29 human colorectal adenocarcinoma cell line. This loss-of-function model allows researchers to interrogate the cellular consequences of abrogating ACOT13-mediated hydrolysis of acyl-CoA thioesters. As a polyclonal pool, these cells provide a heterogeneous genetic background suitable for population-level studies of gene function without requiring single-cell cloning. The product is designed for in vitro investigation of lipid metabolism, signaling pathway modulation, and cancer cell biology.
The HT29 host cell line is an epithelial cell line derived from a human colorectal adenocarcinoma. HT29 cells harbor a mutant p53 gene and are microsatellite stable, making them a well-established model for studying colorectal cancer biology and intestinal epithelial function. Their robust growth characteristics and well-characterized signaling networks render them particularly suitable for metabolic studies, including those examining fatty acid utilization, peroxisomal activity, and energy homeostasis. The mutant p53 background also provides a context for exploring interactions between tumor suppressor pathways and lipid metabolism.
ACOT13 encodes a thioesterase that specifically hydrolyzes medium- and long-chain fatty acyl-CoA molecules, such as palmitoyl-CoA and oleoyl-CoA, to generate free fatty acids and coenzyme A. By controlling intracellular acyl-CoA/CoA ratios, ACOT13 directly influences free fatty acid pools that serve as ligands for peroxisome proliferator-activated receptors (PPARs), including PPARA and PPARG. The enzyme is transcriptionally regulated by PPARA, PPARG, SREBF1, and LXRA, and is responsive to dietary fatty acids. Downstream, ACOT13 activity modulates AMPK signaling and fatty acid oxidation pathways, intersecting with carnitine palmitoyltransferase 1 (CPT1) and acyl-CoA synthetase (ACSL) functions. ACOT13 is known to homodimerize and putatively interacts with thioredoxin, suggesting additional regulatory layers.
In the context of HT29 colorectal cancer cells, disrupting ACOT13 provides a powerful tool to dissect the role of acyl-CoA hydrolysis in tumor cell metabolism. Colorectal cancer cells frequently reprogram lipid metabolism to support proliferation and survival; ACOT13 may contribute by altering the availability of fatty acids for ??-oxidation or signaling. This knockout model enables investigation of how loss of ACOT13 affects PPAR-driven transcriptional programs, energy homeostasis, and sensitivity to metabolic stress. It is particularly relevant for studying links between obesity, dietary fat, and colorectal cancer progression, as ACOT13 sits at the intersection of lipid handling and nuclear receptor signaling.
Researchers can employ these ACOT13 knockout polyclonal cells in a variety of assays, including western blotting and RT-qPCR to confirm gene disruption and assess downstream targets, free fatty acid quantification and CoA measurement to evaluate metabolic changes, lipidomics for comprehensive lipid profiling, and Seahorse metabolic flux analysis to quantify oxidative metabolism. Functional phenotyping can involve cell proliferation and migration assays to determine the impact on tumorigenic behavior, while PPAR reporter assays can directly measure pathway activation. This model is well-suited for studies on cancer metabolic reprogramming, peroxisomal function, and the development of therapeutics targeting metabolic syndrome and type 2 diabetes. For further details or to inquire about custom services, please contact Ascent Research.
