The DNAL1 Knockout Raji Polyclonal Cells product comprises a heterogeneous pool of Raji B lymphocytes engineered via CRISPR/Cas9 to disrupt the DNAL1 gene. This polyclonal knockout population contains a spectrum of editing events at the target locus, providing a representative loss-of-function model without single-cell cloning. The cells are suitable for studying DNAL1-dependent functions in an immortalized B-cell background.
The Raji cell line is an EBV-positive Burkitt’s lymphoma-derived suspension cell line that expresses characteristic B-cell markers including CD19, CD20, and surface immunoglobulin. Widely used as a model for B-cell malignancies and EBV-associated lymphomagenesis, Raji cells provide a robust and experimentally tractable system for gene perturbation studies. As an immortalized line, they support stable expansion and are amenable to various downstream analyses.
DNAL1 encodes dynein axonemal light chain 1, a leucine-rich repeat protein that serves as a structural component of the outer dynein arm within cilia and flagella. DNAL1 directly interacts with other dynein subunits such as DNALI1, DNAI1, and DNAH5, and with docking complex components including CCDC114 and ARMC4. Assembly and regulation of the outer dynein arm are controlled by transcription factors FOXJ1 and RFX family members, with ZMYND10 and MCIDAS acting as critical co-regulators. In motile cilia, DNAL1 function is essential for generating ciliary beat frequency and coordinating mucociliary clearance, while in sperm flagella it contributes to progressive motility. Loss of DNAL1 disrupts these processes, leading to phenotypes characteristic of primary ciliary dyskinesia and Kartagener syndrome, including situs inversus and chronic respiratory infections.
Although Raji cells are non-ciliated, DNAL1 knockout in this background offers a unique opportunity to explore potential non-ciliary functions of dynein arm components, such as roles in intracellular trafficking, cell cycle regulation, or immune cell signaling. Expression profiling and proteomic approaches in this polyclonal knockout can reveal interactions with NME8 and other dynein-related proteins independently of ciliary architecture. The lymphocytic context may uncover previously unrecognized contributions of DNAL1 to B-cell physiology or EBV biology. Because the knockout population retains cellular heterogeneity, it better mimics the genetic variability observed in patient samples compared to monoclonal knockouts.
Typical applications include screening for compounds that modulate dynein arm assembly in a simplified cellular environment, assessing DNAL1 protein stability by Western blotting, quantifying transcript levels via RT-qPCR, and mapping protein?Cprotein interactions by co-immunoprecipitation and mass spectrometry. Flow cytometry can be used to confirm retention of B-cell markers, while proximity ligation assays probe spatial relationships between DNAL1 and its binding partners. Co-culture or heterologous functional assays may measure ciliary beat rescue when wild-type DNAL1 is reintroduced. This knockout product thus serves as a versatile platform for dissecting dynein arm biology and for developing targeted therapies for ciliopathies. For further technical details and lot-specific quality data, please contact Ascent Research.
