The AGK Knockout HT29 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the human HT29 colorectal adenocarcinoma cell line. This product provides a genetically disrupted acyl glycerol kinase (AGK) gene, enabling loss-of-function studies. The polyclonal format supplies a heterogeneous pool of edited cells, suitable for assessing gene function without clonal selection bias, facilitating mechanistic and phenotypic analyses of AGK-dependent signaling in colorectal cancer.
HT29 is a widely used human colorectal adenocarcinoma cell line established from a 44-year-old female. It maintains an epithelial phenotype and is characterized by robust proliferation and tumorigenic potential in xenografts. With well-documented mutations in pathways such as PIK3CA and BRAF, HT29 cells offer a relevant system for investigating colorectal cancer biology, including migration, invasion, and drug response, providing an ideal host for evaluating AGK disruption.
AGK encodes a lipid kinase that phosphorylates monoacylglycerol and diacylglycerol to produce lysophosphatidic acid (LPA) and phosphatidic acid. LPA activates LPA receptors, driving proliferation and migration. AGK also binds and phosphorylates the tumor suppressor DLC1, inhibiting its RhoGAP activity and leading to sustained RhoA/ROCK signaling, cytoskeletal reorganization, and oncogenic phenotypes. Additionally, AGK localizes to mitochondria, interacting with DRP1 and ADP/ATP translocase to modulate lipid metabolism and apoptosis. Upstream regulators include EGF, androgen receptor signaling, and STAT3, while downstream effectors comprise LPA, DLC1, RhoA, ROCK, and YAP/TAZ.
In HT29 colorectal cancer cells, AGK knockout disrupts a pivotal node linking lipid metabolism, cytoskeletal dynamics, and apoptosis. HT29 cells produce and respond to LPA, contributing to malignant properties. AGK loss is expected to reduce LPA generation, attenuating LPA receptor signaling and downstream cascades such as Rho/ROCK and YAP/TAZ. Moreover, impaired mitochondrial function may sensitize cells to apoptosis, allowing dissection of metabolic stress and cell death pathways in a clinically relevant background.
These polyclonal knockout cells support diverse assays, including LPA quantification, western blotting, RT-qPCR, and phospho-signaling analysis to validate pathway disruption. Rho activation assays and Transwell migration/invasion assays probe cytoskeletal and motility changes, while immunofluorescence and apoptosis assays assess mitochondrial morphology and cell death. They are also ideal for drug target validation, enabling compound testing against LPA signaling or Rho/ROCK pathways. For further details, please contact Ascent Research.
