The ANXA1 Knockout HT29 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the HT29 human colorectal adenocarcinoma cell line. This product provides a heterogeneous pool of cells with targeted disruption of the ANXA1 gene, enabling loss-of-function studies without the selection of a single clonal isolate. The polyclonal format preserves the natural genetic diversity of the parental line while eliminating ANXA1 protein expression, offering a robust model for functional genomics and drug screening applications.
The HT29 cell line, originally isolated from a primary colorectal adenocarcinoma, serves as an extensively characterized epithelial model for colorectal cancer. HT29 cells exhibit adherent growth, retain the ability to differentiate into enterocyte-like cells under appropriate culture conditions, and harbor mutations in APC, TP53, and other oncogenic pathways common in colorectal tumors. These features make HT29 cells a widely used system for investigating intestinal epithelial biology, cancer cell signaling, and therapeutic responses.
ANXA1 encodes annexin A1, a glucocorticoid-regulated phospholipid-binding protein that functions as a critical mediator of anti-inflammatory responses. The protein inhibits phospholipase A2, suppresses NF-??B activity, and activates formyl peptide receptor 2 (FPR2/ALX) to promote inflammation resolution. ANXA1 interacts with S100A11, actin, and phospholipids, and is transcriptionally regulated by the glucocorticoid receptor and SP1. Downstream, ANXA1 modulates MAPK signaling, E-cadherin expression, and integrin function, thereby influencing apoptosis, cell proliferation, and membrane trafficking. Loss of ANXA1 disrupts these pathways, leading to enhanced NF-??B activation, altered EGFR signaling, and impaired glucocorticoid responses.
In HT29 colorectal adenocarcinoma cells, ANXA1 knockout mimics the loss-of-function observed in certain colorectal cancer subtypes, where diminished ANXA1 expression correlates with increased tumorigenicity and poor prognosis. This engineered model allows researchers to dissect the tumor-suppressive roles of ANXA1, including its regulation of cell proliferation and apoptosis. Furthermore, the HT29 background provides a clinically relevant platform to study how ANXA1 deficiency affects metastatic potential, inflammatory tumor microenvironment interactions, and resistance to glucocorticoid-based therapies.
Applications include examining ANXA1-dependent modulation of NF-??B and glucocorticoid receptor signaling, assessing cell migration and invasion through Transwell assays, and evaluating proliferation and apoptosis via MTT and annexin V staining. The polyclonal knockout cells are also suitable for drug response profiling, particularly with anti-inflammatory agents or EGFR inhibitors. Additional experimental approaches such as RT-qPCR, western blotting, immunofluorescence, and flow cytometry for FPR2/ALX surface expression can validate mechanistic hypotheses. For further information, please contact Ascent Research.
