The CD19 Knockout AGS Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout population derived from the AGS human gastric adenocarcinoma cell line, designed to disrupt the CD19 gene (Homo sapiens). This pool contains a heterogeneous mix of edited alleles, providing a robust tool for loss-of-function studies without the need for clonal isolation. Although CD19 normally acts as a B cell co-receptor, its targeting in this non-lymphoid epithelial context creates a unique platform for investigating gene knockout outcomes and signaling pathway modulation in a cancer model.
The parental AGS line was established from a poorly differentiated gastric adenocarcinoma and is widely employed as a model for gastric epithelial biology and gastric cancer research. These adherent epithelial cells retain key characteristics of the gastric mucosa, making them suitable for studies of epithelial differentiation, cell cycle progression, and response to therapeutic agents. AGS cells are particularly valuable for examining oncogenic signaling cascades, including those driven by PI3K-Akt and NF-??B, pathways that CD19 is known to modulate in B lymphocytes.
CD19 is a B lymphocyte-specific transmembrane co-receptor that amplifies BCR signaling. Upon BCR activation, Lyn phosphorylates CD19, recruiting PI3K and Vav, which lowers the threshold for Akt, Ras-MAPK, and NF-??B pathways. Upstream regulators include Pax5, Ebf1, and IL-4. CD19 complex includes CD21, CD81, Leu-13, and activates Syk, Btk, PLC??2, driving calcium flux. Since AGS cells lack BCR, this knockout aids CD19 reconstitution studies in epithelial cells.
CD19 knockout in AGS cells is a stringent negative control for CD19-dependent studies due to absent B cell signaling. It helps establish baseline PI3K-Akt and NF-??B activity, aiding CD19 reintroduction experiments. The polyclonal pool is ideal for evaluating CRISPR editing efficiency and off-target effects in a carcinoma setting, reflecting editing heterogeneity of pooled screens. It also permits exploration of non-canonical CD19 functions or expression of disease variants.
Applications include western blotting, RT-qPCR for CD19 loss confirmation, CRISPR cleavage assays (T7E1, Surveyor), and genomic PCR/sequencing for indel analysis. Exogenous CD19 allows immunofluorescence of localization and interactions with CD21, CD81. Polyclonal cells also suit guide RNA optimization and delivery method testing in epithelial cancer models. For protocols and support, contact Ascent Research.
