The DNAAF5 Knockout Raji Polyclonal Cells product consists of a population of Raji B lymphocytes subjected to CRISPR/Cas9-mediated disruption of the DNAAF5 gene, resulting in a heterogeneous polyclonal knockout pool. This format eliminates the need for time-consuming clonal isolation and provides a robust, population-level loss-of-function model. The polyclonal nature reduces the impact of any single off-target event and is well-suited for experiments requiring large cell numbers, such as biochemical assays or high-throughput screens.
The Raji parental cell line is an Epstein-Barr virus (EBV)-positive B lymphocyte line established from a Burkitt??s lymphoma patient. It has been a cornerstone in immunological and cancer research for decades, offering a reliable suspension culture system that recapitulates features of B-cell malignancies. Raji cells are amenable to electroporation and other delivery methods, enabling efficient CRISPR/Cas9 gene editing and subsequent functional analyses, including flow cytometry, co-immunoprecipitation, and protein interaction studies.
DNAAF5 encodes a cytoplasmic co-chaperone essential for the preassembly of axonemal dynein complexes, the multi-subunit motors required for ciliary and flagellar movement. This protein functions downstream of the master transcription factors FOXJ1 and RFX3, which orchestrate ciliary gene expression. DNAAF5 physically associates with other dynein assembly factors, specifically DNAAF1, DNAAF2, and DNAAF3, as well as with dynein intermediate chains DNAI1 and DNAI2, facilitating the proper folding and incorporation of heavy chains such as DNAH5 and DNAH11 into the axonemal dynein complex. Mutations in DNAAF5 lead to defective dynein arm assembly, immotile cilia, and clinical manifestations of primary ciliary dyskinesia, including Kartagener syndrome. Although Raji B lymphocytes are non-ciliated, the expression of DNAAF5 suggests potential extra-ciliary functions that remain to be elucidated, making the knockout model a valuable tool for discovery.
In the Raji B-cell context, disruption of DNAAF5 allows researchers to probe its non-canonical roles in lymphocyte biology and lymphoma pathogenesis. Because the protein??s chaperone activity may extend to non-dynein clients, knockout cells could reveal impacts on protein folding, cell cycle progression, or apoptosis. This model is particularly relevant for investigating how genes traditionally linked to ciliopathies may contribute to hematological malignancy and immune dysregulation, thereby expanding their functional repertoire beyond motile cilia.
This polyclonal knockout pool supports various applications: Western blotting and RT-qPCR for confirming gene disruption, flow cytometry for cell cycle and apoptosis analysis, and co-immunoprecipitation to probe dynein assembly factor interactions. It is also suitable for functional genomics screens and drug response studies. For additional product information or a quote, contact Ascent Research.
