The P4HA2 Knockout Raji Polyclonal Cells are a CRISPR/Cas9-edited polyclonal population originating from the Raji B lymphocyte line, engineered for disruption of the P4HA2 gene. This loss-of-function model enables investigation of prolyl 4-hydroxylase subunit alpha 2 in collagen modification and extracellular matrix (ECM) dynamics. The polyclonal format maintains genetic diversity, facilitating bulk functional studies suitable for diverse downstream assays without the need for clonal isolation.
Raji is a human Burkitt’s lymphoma-derived B lymphocyte line, Epstein-Barr virus positive, extensively used in immunology for antibody production and antigen presentation studies. As a suspension cell line, it allows scalable culture and high-throughput phenotypic screening. Its malignant phenotype provides a relevant context for B-cell lymphoma research, particularly for exploring tumor-microenvironment crosstalk and adaptive immune processes.
P4HA2 encodes the catalytic alpha-subunit of collagen prolyl 4-hydroxylase, which complexes with P4HB to hydroxylate proline residues in procollagens such as COL1A1, COL1A2, and COL3A1, essential for triple-helix stability and ECM integrity. Expression is upregulated by HIF1A under hypoxia and is responsive to TGFB1 and EGF. The HIF1A-P4HA2-P4HB-COL1A1 signaling axis integrates oxygen sensing with matrix biosynthesis, while TGFBR-mediated pathways further modulate fibrotic and ECM remodeling responses.
In the Raji background, P4HA2 knockout disrupts collagen hydroxylation, likely impairing cell adhesion, migration, and ECM-dependent signaling within the lymphoma niche. This alteration provides a platform to dissect how collagen modifications influence B-cell lymphoma progression and interactions with the tumor stroma, where hypoxia and TGF-beta cues are critical.
Applications include studying collagen modification in B-cell lymphoma, ECM roles in the tumor microenvironment, and hypoxia response mechanisms. The cells are suitable for screening P4HA2 inhibitors and evaluating tumor-stromal interactions. Assays commonly employed are western blotting for hydroxylated collagen, RT-qPCR, hydroxyproline-based collagen synthesis measurement, migration/invasion assays, ECM immunofluorescence, RNA-seq, metabolic analysis under hypoxia, and flow cytometry for integrins. For additional details, please contact Ascent Research.
