The PHIP Knockout Raji Polyclonal Cells constitute a polyclonal knockout cell population engineered via CRISPR/Cas9-mediated gene disruption of the PHIP locus in the Raji B lymphocyte line. This heterogeneous loss-of-function model captures diverse editing outcomes across the polyclonal pool, allowing phenotypic analysis without clonal isolation. It is suitable for studying PHIP-dependent signaling, cytoskeletal dynamics, and ubiquitin ligase functions in a lymphoblastoid context.
The Raji cell line is a human Burkitt lymphoma-derived B lymphoblast, maintained in suspension culture and characterized by EBV positivity. It recapitulates key features of mature B cells, including surface immunoglobulin expression and responsiveness to cytokine and antigen receptor stimulation. Widely adopted for B cell biology, EBV research, and lymphoma studies, its genetic tractability makes it an ideal host for targeted gene knockout.
PHIP acts as a pleckstrin homology domain-interacting scaffold, bridging insulin receptor (INSR) and IRS1 to actin cytoskeleton reorganization through filamin A binding, thereby promoting GLUT4 translocation and cell migration. PHIP also functions as a substrate receptor (DCAF14) for the CUL4A/B-DDB1 ubiquitin ligase, directing protein targets for proteasomal degradation. Representative signaling nodes include PI3K, AKT, and downstream actin regulators, positioning PHIP at the intersection of metabolic and migratory pathways.
In the Raji B lymphocyte background, PHIP ablation enables dissection of its contributions to cell adhesion, motility, and antigen presentation, processes relevant to lymphomagenesis and immune cell trafficking. Given PHIP??s association with neurodevelopmental disorders and breast cancer, this model facilitates investigation of B-cell-intrinsic roles and potential oncogenic mechanisms. The EBV-positive context further permits study of viral interactions with host insulin/cytoskeletal networks.
These cells are applicable to a broad array of functional assays. Western blotting and RT-qPCR confirm PHIP knockout. Insulin stimulation combined with phospho-AKT flow cytometry evaluates signaling integrity. Co-immunoprecipitation probes PHIP interactions with IRS1, filamin A, or DDB1. Transwell migration assays and phalloidin staining assess motility and actin architecture. Ubiquitination assays and RNA-seq capture alterations in protein degradation and transcriptomic landscapes. The model also supports drug sensitivity screening for PHIP-related therapeutic targets. For detailed technical specifications, please contact Ascent Research.
