The ACSL1 Knockout HT29 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout population targeting the ACSL1 gene in human HT29 colorectal adenocarcinoma cells. This loss-of-function model permits investigation of ACSL1-dependent long-chain fatty acid activation, a critical step in lipid metabolism. The heterogeneous gene-disrupted pool enables functional studies without clonal selection artifacts, supporting dissection of pathways relying on acyl-CoA formation, including triglyceride synthesis, ??-oxidation, and lipid droplet biogenesis.
The HT29 cell line, derived from a 44-year-old Caucasian female with colorectal adenocarcinoma, is a widely used model for intestinal epithelial biology and colorectal cancer. Exhibiting classical adenocarcinoma features, these adherent cells are employed to study barrier function, mucin production, and metabolic reprogramming. Their robust in vitro growth and genetic tractability make them ideal for knockout generation and downstream phenotypic assays. This clinically relevant system is valuable for examining how lipid metabolism influences colorectal tumorigenesis and drug responses.
ACSL1 encodes an acyl-CoA synthetase that catalyzes the ATP-dependent ligation of long-chain fatty acids (C16?CC20) to coenzyme A, generating acyl-CoA thioesters essential for both anabolic and catabolic lipid pathways. Its expression is regulated by PPAR??, SREBP1, LXR, and insulin signaling, responding to fatty acid availability. Acyl-CoA products feed into triglyceride synthesis, mitochondrial ??-oxidation via CPT1, lipid droplet biogenesis, and cholesterol esterification by ACAT1. ACSL1 interacts with FATP1 and acyl-CoA-binding proteins that facilitate intracellular fatty acid trafficking. Thus, ACSL1 serves as a gatekeeper partitioning fatty acids between storage and oxidation, governing cellular energy homeostasis.
In HT29 colorectal adenocarcinoma cells, ACSL1 knockout is a powerful tool for studying cancer metabolic plasticity. Colorectal tumors often exploit lipid-rich microenvironments; HT29 cells upregulate fatty acid metabolism to support proliferation. ACSL1 disruption impairs conversion of exogenous fatty acids to acyl-CoA, reducing triglyceride accumulation, ??-oxidation, and ATP production, particularly under lipid supplementation. This makes the model relevant for exploring how lipid utilization drives tumor growth, metabolic adaptation, and obesity-linked colorectal cancer. It enables investigation of connections between lipid metabolism and oncogenic signaling in malignancy.
The ACSL1 Knockout HT29 Polyclonal Cells are suitable for colorectal cancer metabolism research, obesity and type 2 diabetes modeling, and drug screening targeting fatty acid utilization. Key assays include BODIPY staining for lipid droplets, radiolabeled fatty acid oxidation, triglyceride quantification, and cell proliferation under lipid supplementation. Additional readouts encompass ATP luminescence, western blotting for ACSL1 and CPT1, RT-qPCR for metabolic genes, and Seahorse metabolic flux analysis. These tools allow comprehensive profiling of the metabolic impacts of ACSL1 loss. For product inquiries, contact Ascent Research.
