The CNPY3 Knockout Raji Polyclonal Cells constitute a CRISPR/Cas9-mediated polyclonal knockout cell population derived from the Raji B lymphocyte line. Using CRISPR/Cas9 genome editing, the CNPY3 gene is disrupted to generate a heterogeneous pool of edited alleles across the cell population, providing a loss-of-function model without clonal selection. This polyclonal format enables pooled functional genomic studies and is supplied as living cells ready for expansion in culture.
The Raji cell line is an Epstein-Barr virus (EBV)-positive B lymphocyte line originally established from a Burkitt lymphoma patient. As a lymphoblastoid line, Raji cells retain mature B cell characteristics and are widely employed for investigations of B cell biology, immunoglobulin studies, and lymphoma research. Their rapid growth and stable genetic background make them a robust host for gene-editing applications. In lymphoma contexts, Raji cells exhibit deregulated signaling pathways that contribute to uncontrolled proliferation and survival, rendering them particularly relevant for studying oncogenic mechanisms through targeted gene disruptions.
CNPY3 encodes canopy FGF signaling regulator 3, a positive regulator of fibroblast growth factor (FGF) signaling that facilitates activation of the FGF receptor FGFR1. Mechanistically, CNPY3 interacts with FGFR1 and enhances downstream signal transduction through the MAPK/ERK and PI3K/AKT cascades. Upon FGF ligand binding (e.g., FGF1 or FGF2), FGFR1 recruits adaptor proteins such as FRS2 and GRB2, leading to activation of the RAS?CRAF1?CMAP2K1?CMAPK1/3 (ERK1/2) pathway and the AKT1?CMTOR axis. The downstream effector CCND1 (Cyclin D1) integrates mitogenic signals to drive G1/S cell cycle progression. Hence, CNPY3 functions upstream of these proliferative and survival signals, positioning it as a key modulator of FGF-dependent cellular outcomes.
Disruption of CNPY3 in Raji B lymphocytes offers a valuable model to dissect FGF signaling contributions to lymphoma cell biology. In Burkitt lymphoma cells, aberrant activation of growth factor pathways can promote oncogenesis; therefore, CNPY3 knockout may attenuate FGF-mediated proliferation and survival, potentially sensitizing cells to apoptotic stimuli. This model enables examination of how loss of CNPY3 impacts the activation state of MAPK1/3 and AKT1, alters Cyclin D1 expression, and influences B cell malignancies. Moreover, the polyclonal nature of the knockout population mimics heterogeneous tumor environments more realistically than clonal lines, facilitating studies that require pooled genetic screening or functional complementation analysis.
Typical experimental applications include western blotting to assess changes in total and phosphorylated ERK1/2, AKT, and MTOR levels; cell proliferation assays to quantify growth defects; apoptosis assays to measure sensitivity to death signals; and flow cytometry to evaluate cell cycle distribution or apoptotic markers. RT-qPCR can validate CNPY3 transcript disruption and monitor downstream gene expression changes. Phospho-signaling analysis and inhibitor combination studies allow dissection of pathway crosstalk. Additionally, this knockout model can be used for functional rescue experiments or co-culture systems to investigate interactions within the tumor microenvironment. For further details or to discuss custom requirements, please contact Ascent Research.
