CFAP36 Knockout Raji Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the Raji B lymphoblastoid cell line, designed to disrupt expression of the cilia- and flagella-associated protein CFAP36. This polyclonal model reduces clonal selection artifacts and provides a population-level loss-of-function system ideal for investigating CFAP36??s non-ciliary functions in a lymphoid background.
The Raji host cell line is an Epstein-Barr virus-positive B lymphoblastoid cell line isolated from a Burkitt??s lymphoma patient. These suspension-adapted cells are extensively employed in B cell biology and lymphomagenesis research due to their transformed phenotype and active cycling. Importantly, Raji cells lack primary cilia, creating an optimal cellular context to dissect non-ciliary roles of cilia-associated proteins such as CFAP36.
CFAP36 encodes a protein critical for axonemal assembly and dynein arm formation in motile cilia and flagella, where it participates in microtubule organization. It interacts with core ciliogenic components including SPEF2, IFT88, and IFT20, and associates directly with dynein arms and tubulin within the axoneme. In ciliated cells, CFAP36 expression is governed by FOXJ1 and RFX transcription factors, central regulators of motile ciliogenesis. However, CFAP36??s function in non-ciliated B lymphocytes remains unexplored. Given its connection to microtubule dynamics, CFAP36 may play roles in mitotic spindle assembly, interphase microtubule stability, or intracellular transport in dividing B cells. The CRISPR-mediated knockout in Raji cells allows functional dissection of these potential non-ciliary activities.
By targeting CFAP36 in Raji cells, which naturally lack primary cilia, this model directly probes microtubule-dependent processes essential for lymphoma cell proliferation. Altered cell cycle progression, spindle abnormalities, or changes in genomic stability can be assessed, shedding light on CFAP36??s contribution to B cell malignancy. Moreover, because CFAP36 mutations are linked to ciliopathies like primary ciliary dyskinesia and situs inversus, this knockout provides a platform to understand tissue-specific consequences of CFAP36 deficiency outside ciliated tissues. The polyclonal population ensures biological variability is maintained, enhancing reproducibility in functional studies and drug sensitivity testing.
Research applications include investigating CFAP36??s role in microtubule organization and division in B cells, using assays such as immunofluorescence for spindle morphology, flow cytometry for cell cycle analysis, and proliferation/apoptosis assays. Transcriptomic changes can be profiled via RNA-seq, and protein-level disruption verified by Western blotting and RT-qPCR. The cells are also valuable for drug target validation, particularly evaluating sensitivity to microtubule-targeting chemotherapeutics. For further details, please contact Ascent Research.
