The FIG4 Knockout Raji Polyclonal Cells are a CRISPR/Cas9-edited polyclonal cell population derived from Raji B lymphocytes, designed to eliminate FIG4 gene function. Unlike single-cell clones, this polyclonal pool captures population-level heterogeneity while ensuring robust target-gene disruption, making it suitable for high-throughput functional genomics and biochemical assays. The cells are provided as a proliferating culture and can be used directly for validation via Western blotting or RT-qPCR.
The Raji host cell line is an EBV-positive Burkitt lymphoma line widely employed in immunology and cancer research. Its B lymphocyte origin and rapid suspension growth facilitate studies of B cell receptor signaling, viral oncogenesis, and lymphomagenesis. The line’s established genetic tractability and compatibility with multi-well formats support efficient integration into drug screening and pathway analysis workflows.
FIG4 encodes a 5-phosphatase that specifically dephosphorylates PtdIns(3,5)P2 to produce PtdIns3P, a critical step in endolysosomal phosphoinositide metabolism. This reaction occurs within the PIKFYVE-FIG4-VAC14 regulatory complex, where VAC14 scaffolds the assembly and PIKFYVE synthesizes the substrate. The resulting PtdIns3P pool governs endolysosomal maturation, autophagic clearance, and lysosomal ion homeostasis via effectors such as TRPML1 and TFEB. FIG4 also influences mTORC1 signaling, connecting lipid signaling to nutrient-dependent growth control. Disruption of FIG4 thus profoundly impacts cellular homeostasis.
In the Raji B lymphocyte context, FIG4 knockout perturbs endolysosomal trafficking and autophagy, processes that may affect antigen presentation and survival signaling in lymphoma. This model enables investigation of how lysosomal dysfunction contributes to drug resistance or synthetic lethality in B cell malignancies. Additionally, given FIG4 mutations in Charcot-Marie-Tooth type 4J, amyotrophic lateral sclerosis, and Yunis-Varon syndrome, the cells offer a unique platform to study neurodegeneration-linked pathways in a cancer setting, exploring cross-disease mechanisms of lysosomal stress.
Typical applications include monitoring autophagy flux via LC3 immunofluorescence or flow cytometry, assessing lysosomal pH with ratiometric dyes, and performing drug sensitivity assays with chemotherapeutics. Western blotting for FIG4 or downstream targets like LAMP2 can confirm knockout and pathway alterations. The polyclonal nature also supports co-culture and in vivo xenograft studies to evaluate tumor microenvironment interactions. For further inquiries or to discuss specialized applications, please contact Ascent Research.
