The BRAF Knockout AGS Polyclonal Cells product is a CRISPR/Cas9-edited polyclonal human AGS cell population with targeted disruption of the BRAF gene. This polyclonal knockout pool provides a genetically heterogeneous loss-of-function model, avoiding clonal biases and enabling unbiased functional studies of BRAF-dependent signaling, pathway dependency, and drug sensitivity profiling.
The parental AGS cell line, derived from a human primary gastric adenocarcinoma, is an adherent epithelial line widely used to model gastric cancer. AGS cells retain gastric epithelial characteristics, support Helicobacter pylori infection, and are responsive to chemotherapeutic agents, making them a robust platform for dissecting oncogenic signaling and host-pathogen interactions.
BRAF encodes a serine/threonine kinase that transmits signals from RAS GTPases to MEK1/2 in the MAPK/ERK pathway. Activated upstream by KRAS, NRAS, HRAS, and receptor tyrosine kinases such as EGFR and PDGFR with input from SRC kinases, BRAF phosphorylates MEK1 (MAP2K1) and MEK2 (MAP2K2), which then activate ERK1 and ERK2. This cascade regulates transcription factors including ELK1 and MYC. BRAF activity is modulated by interactions with 14-3-3 proteins, HSP90, CRAF (RAF1), and KSR scaffold proteins. Oncogenic BRAF mutations like V600E result in constitutive kinase activity and drive tumorigenesis.
In AGS gastric adenocarcinoma cells, BRAF knockout enables precise dissection of MAPK pathway dependency. Although BRAF mutations are less common in gastric cancer, pathway hyperactivation frequently occurs via upstream receptor overexpression or RAS alterations. This model facilitates the identification of compensatory signaling mechanisms, such as CRAF-driven bypass, and resistance to RAF and MEK inhibitors. It also provides a system to investigate how H. pylori infection intersects with host cell MAPK signaling to modulate oncogenic processes.
Researchers can apply this knockout pool in western blot analyses of phospho-MEK and phospho-ERK, RT-qPCR for BRAF transcript, and MTT or Annexin V assays to assess proliferation and apoptosis. Transwell invasion and migration assays, drug sensitivity testing with vemurafenib, and flow cytometric cell cycle analysis further extend its utility, along with RNA-seq transcriptome profiling. The model is suited for gastric adenocarcinoma tumor biology studies and host-pathogen interaction experiments with H. pylori. For further information, please contact Ascent Research.
