The DDAH1 Knockout Raji Polyclonal Cells are a CRISPR/Cas9-edited polyclonal population engineered to disrupt the DDAH1 gene in Raji B lymphocytes. This polyclonal format maintains population heterogeneity and provides a stable loss-of-function model for interrogating the ADMA/nitric oxide signaling axis in B-cell lymphoma, avoiding clonal selection artifacts.
Raji cells are an Epstein-Barr virus-positive B lymphocyte line derived from a Burkitt’s lymphoma patient. Widely employed in immunology and cancer biology, they exhibit robust proliferation and are well-characterized for studying lymphomagenesis, immune synapse dynamics, and drug sensitivity. Their high transfectability renders them an ideal host for CRISPR-based genomic perturbation.
DDAH1 hydrolyzes asymmetric dimethylarginine (ADMA) and N-monomethyl-L-arginine (L-NMMA), both endogenous inhibitors of nitric oxide synthase (NOS) isoforms. By lowering cellular ADMA, DDAH1 relieves NOS inhibition, increasing nitric oxide (NO) production that stimulates soluble guanylyl cyclase, cGMP, and protein kinase G (PKG). DDAH1 activity is modulated by TNF-??, VEGF, shear stress, reactive oxygen species, and KLF2, and the enzyme physically interacts with tubulin while functionally coupling to protein arginine methyltransferases (PRMTs) and DDAH2.
In the Raji B-cell context, DDAH1 knockout enables dissection of how the ADMA?CNO pathway influences lymphoma cell behavior, including proliferation, apoptosis, migration, and angiogenesis. It may reveal crosstalk between NO signaling and B-cell receptor or Epstein-Barr virus latency programs, shedding light on mechanisms of lymphomagenesis and potential therapeutic targets.
Representative applications encompass Western blotting for DDAH1 and phospho-eNOS, RT-qPCR profiling of DDAH1, NOS isoforms, and arginine-metabolism genes, Griess and DAF-FM assays for NO quantification, and ADMA measurement by ELISA or LC-MS. Functional studies can employ MTS/BrdU proliferation assays, Annexin V flow cytometry for apoptosis, transwell migration assays, and drug sensitivity testing with NOS inhibitors such as L-NAME. These cells also support co-immunoprecipitation for interaction partners and transcriptome-wide RNA-seq analysis. For further details, please contact Ascent Research.
