The ASL Knockout HT29 Polyclonal Cells product is a CRISPR/Cas9-edited polyclonal knockout population derived from the human HT29 colorectal adenocarcinoma line, featuring disruption of the ASL gene. This model enables loss-of-function studies of argininosuccinate lyase (ASL) within a colon cancer cell context. The polyclonal composition preserves the heterogeneous genetic landscape of the HT29 line, allowing functional assessment of ASL disruption across a range of editing outcomes. Supplied as live cells, it is intended for research in cancer metabolism, urea cycle biology, and arginine signaling.
The HT29 host cell line originated from a primary colorectal adenocarcinoma of a 44-year-old Caucasian female, displaying adherent epithelial morphology and harboring oncogenic mutations in APC, p53, and KRAS. These characteristics make it a representative model for studying intestinal differentiation, drug transport, and colorectal cancer biology, with extensive use in investigating colonocyte barrier function, metabolic reprogramming, and colorectal cancer progression.
ASL encodes argininosuccinate lyase, which catalyzes the cleavage of argininosuccinate into arginine and fumarate, the final step of the urea cycle, and provides substrates for nitric oxide (NO) and polyamine synthesis. Regulated by HNF-4??, glucocorticoid receptor signaling, CREB, and PPAR??, ASL interacts with ASS1, NOS, and arginase within the urea cycle pathway that includes CPS1, OTC, ASS1, and ARG1. ASL disruption in HT29 cells is predicted to reduce arginine and fumarate, altering NO signaling, polyamine synthesis, and fumarate-dependent pathways that influence proliferation and survival.
In the HT29 background with mutant APC, p53, and KRAS, ASL knockout facilitates investigation of urea cycle defects in tumor metabolism and arginine auxotrophy, where cells rely on exogenous arginine. The model enables dissection of fumarate-mediated signaling and its intersection with oncogenic pathways, and is relevant for studying argininosuccinic aciduria and the role of mitochondrial urea cycle enzymes in colonocyte biology.
Applications include RT-qPCR and western blotting for ASL expression, argininosuccinate lyase activity assays, mass spectrometric quantification of arginine and fumarate, NO detection by Griess assay, and cell viability testing under arginine deprivation. Immunofluorescence for enzyme localization and drug screening for ASL deficiency therapies further extend utility. For technical support, contact Ascent Research.
