The ANXA3 Knockout HT29 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population generated from the HT29 human colon carcinoma cell line, designed for loss-of-function investigation of the ANXA3 gene. This heterogeneous polyclonal pool disrupts ANXA3 expression without clonal selection, minimizing artifacts and better reflecting the genetic diversity of tumor cell populations. Researchers can use this model to study ANXA3-dependent phenotypes while avoiding biases introduced by single-cell cloning.
The HT29 parental line is an epithelial cell model derived from a primary colorectal adenocarcinoma of a 44-year-old Caucasian female. It is widely employed to study intestinal epithelial differentiation, barrier function, and mucus production. Under conducive culture conditions, HT29 cells form polarized monolayers with tight junctions, making them particularly valuable for examining epithelial-mesenchymal transition (EMT) and tumor microenvironment interactions in colorectal cancer.
ANXA3 encodes Annexin A3, a calcium-dependent phospholipid-binding protein that coordinates membrane organization, exocytosis, vesicle trafficking, cell proliferation, migration, and apoptosis. Its expression is regulated by transcription factors SP1, NF-??B, and AP-1, and is induced by EGF and TNF-??. ANXA3 interacts with S100A4, annexin A2, actin, tubulin, and the PI3K catalytic subunit, thereby modulating cytoskeletal dynamics and signal transduction. ANXA3 activates MAPK/ERK and PI3K/AKT pathways, leading to phosphorylation of ERK and AKT and upregulation of downstream targets such as cyclin D1 (CCND1), Bcl-2 (BCL2), and MMP9, which collectively promote proliferation, survival, and invasiveness.
In the HT29 colon carcinoma context, ANXA3 knockout disrupts calcium-dependent membrane dynamics and cytoskeletal reorganization, resulting in diminished cell proliferation and migration. These effects are mediated through downregulation of MAPK/ERK and PI3K/AKT signaling, as evidenced by reduced phospho-ERK and phospho-AKT levels, decreased cyclin D1 and Bcl-2 expression, and enhanced apoptosis. This phenotype is directly relevant to colorectal cancer progression, where ANXA3 overexpression is associated with tumor aggressiveness, EMT, and chemoresistance, making the knockout cells a powerful isogenic system for mechanistic studies.
This polyclonal knockout model is suitable for diverse applications, including colorectal cancer progression studies, EMT modeling, drug resistance research, and anti-cancer drug screening. Representative assays include western blotting and RT-qPCR for expression profiling, Transwell migration/invasion assays, annexin V/PI apoptosis detection, flow cytometry for cell cycle analysis, and MTT/CCK-8 drug sensitivity testing. Phospho-ERK/AKT ELISAs provide precise quantification of pathway activation. For additional information, please contact Ascent Research.
