The AGTRAP Knockout HT29 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the human colorectal adenocarcinoma cell line HT29. This product provides a heterogeneous pool of cells harboring CRISPR/Cas9-mediated disruption of the AGTRAP gene, enabling loss-of-function studies without clonal selection. The polyclonal format preserves genetic diversity inherent to the knockout pool, avoiding artifacts potentially introduced by single-cell cloning, and is well-suited for population-level signaling and phenotypic analyses.
The HT29 cell line is a widely utilized model of intestinal epithelial biology and colorectal cancer. Established from a primary human colorectal adenocarcinoma, HT29 cells exhibit adherent epithelial morphology and express carcinoembryonic antigen (CEA) and the intestinal transcription factor CDX2. These cells are competent in angiotensin II type 1 receptor (AT1R) signaling, making them an appropriate host for investigating AGTRAP function within a colorectal cancer context.
AGTRAP (Angiotensin II Receptor-Associated Protein) functions as a negative regulator of AT1R signaling. It interacts with the carboxyl-terminal tail of AT1R, facilitating receptor internalization and desensitization through interactions with ??-arrestin and G protein-coupled receptor kinases (GRKs). Physiologically, AGTRAP modulates the renin-angiotensin system and blood pressure. Downstream of AT1R, AGTRAP attenuates activation of extracellular signal-regulated kinase 1/2 (ERK1/2), signal transducer and activator of transcription 3 (STAT3), nuclear factor kappa-light-chain-enhancer of activated B cells (NF-??B), phospholipase C beta (PLC??), and calcium/calmodulin pathways. Consequently, AGTRAP disruption via CRISPR/Cas9 results in enhanced AT1R-mediated MAPK/ERK and STAT3 signaling upon angiotensin II stimulation.
In HT29 cells, which endogenously express AT1R and downstream effectors, AGTRAP knockout creates a hyper-responsive angiotensin II signaling environment. This model is highly relevant for dissecting the role of angiotensin II signaling in colorectal cancer progression, where aberrant activation of MAPK/ERK and STAT3 pathways contributes to proliferation, survival, and inflammation. Moreover, it allows the exploration of crosstalk between the renin-angiotensin system and oncogenic signaling networks, potentially revealing therapeutic vulnerabilities in AT1R-expressing colorectal tumors.
Researchers can employ this knockout model in diverse experimental workflows, including Western blotting for AGTRAP and phospho-ERK1/2, RT-qPCR verification of AGTRAP mRNA depletion, calcium mobilization assays, and cell proliferation studies following angiotensin II challenge. Additional applications encompass GPCR regulation studies, drug screening for AT1R modulators, and reporter assays for STAT3 and NF-??B activity. These cells are a valuable tool for preclinical investigations into cardiovascular disease and colorectal cancer. For further details or custom requests, please contact Ascent Research.
