CRTAP Knockout Raji Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout population targeting the CRTAP gene in the Raji B lymphocyte line. Supplied as a heterogeneous pool of CRTAP-disrupted cells, this model enables loss-of-function studies of collagen prolyl 3-hydroxylation without clone isolation, facilitating immediate use in extracellular matrix biology and skeletal dysplasia research. The polyclonal nature ensures representation of multiple disruption events, providing a robust system for analyzing collagen biosynthesis, ER stress, and cell-matrix interactions.
The host Raji cell line originates from an EBV-immortalized B lymphocyte of a Burkitt lymphoma patient, displaying a B-cell phenotype suited for immune defense, antibody secretion, and antigen presentation studies. Although not a primary collagen producer, Raji cells express ECM components and collagen receptors, making them relevant for investigating cell-matrix adhesion and migration. Their continuous proliferation supports reproducible assays and transgenic manipulations.
CRTAP is a subunit of the collagen prolyl 3-hydroxylase complex, together with P3H1 (LEPRE1) and cyclophilin B (PPIB), which catalyzes 3?hydroxylation of prolines in COL1A1 and COL1A2 procollagens, essential for triple-helix folding and secretion. Regulated by TGF???/BMP pathways and transcription factors SP1 and NF-??B, CRTAP disruption destabilizes the complex, leading to accumulation of misfolded collagen in the ER and activation of ER stress responses involving chaperones such as HSP47.
In Raji B cells, CRTAP knockout impairs secretion of collagen isoforms, potentially disrupting integrin-mediated adhesion and immune synapse organization. The resulting ER stress from misfolded collagen accumulation offers a model to study proteostatic imbalance in professional secretory cells, linking collagenopathy defects to hematopoietic cell function. This provides a unique opportunity to dissect cell-autonomous effects of collagen mutations on B-cell biology, including antigen presentation and antibody secretion dynamics.
Applications include western blotting and RT-qPCR for CRTAP, COL1A1/COL1A2, and ER stress markers; immunofluorescence for collagen localization; collagen secretion assays; cell adhesion/migration assays; and ER stress reporter screens for osteogenesis imperfecta type VII drug discovery. These assays enable detailed mechanistic studies of collagen biosynthesis, extracellular matrix organization, and therapeutic targeting of collagenopathies. For further assistance, please contact Ascent Research.
