The OFD1 Knockout Raji Polyclonal Cells are a CRISPR/Cas9-engineered polyclonal knockout population targeting the OFD1 gene in human Raji B lymphocytes. The polyclonal pool contains a spectrum of loss-of-function alleles, obviating the need for single-cell cloning and providing a genetically diverse model for pooled functional studies. This format is ideal for biochemical and cell-based assays requiring population-level readouts.
Derived from an EBV-positive Burkitt lymphoma, Raji cells are a prototypical B lymphocyte line capable of antibody production and antigen presentation. Their robust growth, stable karyotype, and well-characterized oncogenic signaling pathways make them a versatile host for gene editing. The EBV background further permits exploration of viral latency and host gene interplay, while their B-cell attributes facilitate studies of humoral immunity and lymphomagenesis.
OFD1 is a centrosomal satellite scaffold critical for primary cilia assembly and Hedgehog signaling. It forms complexes with CEP290, PCM1, centrin, LCA5, and SDCCAG8, coordinating centriolar duplication and protein trafficking. Downstream, OFD1 regulates GLI1?C3 transcription factors via the SHH-PTCH1-SMO cascade, modulated by SUFU and KIF7. Upstream regulators include E2F transcription factors and the MTOR-ULK1 autophagy pathway, linking ciliogenesis to metabolic cues. Disruption of OFD1 abolishes cilium formation and dampens Hedgehog target gene expression.
Although Raji cells lack a primary cilium, OFD1 exerts non-ciliary functions in centriole dynamics and autophagy. In this lymphoma setting, knockout perturbs Hedgehog/GLI transcriptional output and cell cycle progression, relevant to B-cell malignant transformation. This model thus bridges ciliopathy and cancer research, enabling dissection of OFD1??s roles in immune cell proliferation and signaling without the confounding influence of ciliary structures. Additionally, it supports exploration of centrosomal protein contributions to antibody production and immune responses.
Key applications include immunofluorescence analysis of centrosomal markers, western blotting for OFD1 and Hedgehog components, and RT-qPCR of GLI targets. Functional assays such as cell cycle analysis, apoptosis assays, and migration tests are well-suited to the polyclonal model. Drug screening for ciliopathy or autophagy modulators, as well as genetic modifier screens, can leverage the population-level heterogeneity. For further information or assay customization, please contact Ascent Research.
