The DYNC2H1 Knockout Raji Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from Raji B lymphocytes, designed to disrupt the DYNC2H1 gene. This heterogeneous pool provides a loss-of-function model without clonal selection, suitable for investigating DYNC2H1-dependent processes. The mixed genotype recapitulates biological variability, enhancing physiological relevance. The polyclonal format allows study of ciliogenesis and Hedgehog signaling, which are inducible in this lymphoblastoid line, making it a versatile tool for functional genomics.
Raji cells are an immortalized human B lymphocyte line from an EBV-positive Burkitt lymphoma, widely used in immunology and oncology. They retain antibody production and antigen presentation capabilities. Notably, these hematopoietic cells can be induced to form primary cilia upon serum starvation, enabling ciliary studies in an immune context. This unique feature allows exploration of dynein-2 function in B lymphocytes and the impact of gene disruption on ciliary dynamics.
DYNC2H1 encodes the heavy chain of cytoplasmic dynein-2, which drives retrograde intraflagellar transport (IFT) in primary cilia. The motor complex includes DYNC2LI1, WDR34, and WDR60, and interacts with IFT43 and IFT122. Its expression is regulated by RFX transcription factors and FoxJ1. Retrograde IFT is essential for Hedgehog signal transduction; upon pathway activation, SMO accumulation in cilia promotes processing of GLI2 and GLI3 transcription factors, with SUFU as a negative regulator. DYNC2H1 disruption impairs ciliogenesis and Hedgehog signaling, relevant to ciliopathies like short-rib polydactyly syndrome and Jeune asphyxiating thoracic dystrophy.
In the Raji B lymphocyte context, the DYNC2H1 knockout enables study of ciliary and non-ciliary dynein-2 functions. Hedgehog signaling influences B cell development and lymphomagenesis, so DYNC2H1 loss may affect cell proliferation or immune functions. Potential roles in organelle transport can also be examined. This model bridges ciliopathy research and immune cell biology.
These cells support ciliogenesis assays via serum starvation and immunofluorescence for ciliary markers (acetylated ??-tubulin, ARL13B). Hedgehog activity can be measured by GLI1 RT-qPCR or GLI-luciferase reporter. Western blotting and co-immunoprecipitation reveal protein interactions. The polyclonal format enables pooled drug screens for ciliopathy therapy development. This model is valuable for academic and pharmaceutical researchers studying ciliopathy mechanisms and developing therapies. For more information, contact Ascent Research.
