The ACY1 Knockout HT29 Polyclonal Cells provide a CRISPR/Cas9-edited polyclonal knockout cell population for investigating aminoacylase 1 (ACY1) function in a colorectal adenocarcinoma model. This loss-of-function pool is generated by CRISPR/Cas9-mediated disruption of the ACY1 gene in HT29 cells, yielding a heterogeneous knockout population suitable for population-level phenotypic and biochemical analyses without clonal selection bias. This product supports robust functional assays to probe ACY1-dependent processes in cancer metabolism.
The HT29 host line originates from a human female colorectal adenocarcinoma and serves as a well-characterized intestinal epithelial cell model. These cells carry BRAF V600E, TP53, and PIK3CA mutations and are microsatellite stable, representing a common genetic landscape of colorectal cancer. Their adherent growth and well-characterized signaling networks facilitate diverse experimental approaches, enabling studies of metabolic pathway contributions to tumorigenesis and drug sensitivity.
ACY1 encodes a zinc-dependent homodimeric enzyme that hydrolyzes N-acetylated amino acids (e.g., N-acetylhistidine, N-acetyltyrosine) into free amino acids and acetate, essential for amino acid recycling. ACY1??s role in recycling N-acetylated amino acids is crucial for maintaining intracellular amino acid homeostasis. The enzyme??s activity is governed by transcriptional regulators of amino acid metabolism and in turn influences mTOR signaling through modulation of amino acid pools. Its interacting factors include homodimerization and Zn2+ cofactor, and it occupies a key node linking amino acid catabolism to metabolic signaling.
In HT29 cells, ACY1 knockout impairs N-acetylated amino acid hydrolysis, leading to substrate accumulation and reduced flux into free amino acids and acetate. This metabolic disruption likely dysregulates mTOR signaling, a pathway hyperactivated by PIK3CA mutation, potentially altering proliferation and survival. The model thus enables dissection of how amino acid metabolism interfaces with oncogenic signaling in BRAF/TP53 mutant colorectal cancer, potentially revealing metabolic vulnerabilities.
Applications include metabolic profiling via LC-MS or HPLC to quantify acetylated amino acids and acetate, coupled with western blotting and RT-qPCR for pathway analysis. Cell proliferation, apoptosis, and migration assays can assess phenotypic outcomes, while metabolomics reveals global metabolic rewiring. This polyclonal knockout population serves as a robust tool for functional studies of amino acid metabolism in cancer and for drug sensitivity screens targeting these pathways. This model is particularly suited for investigating metabolic adaptations in colorectal tumors and identifying new therapeutic targets. For further information, please contact Ascent Research.
