The DNTTIP1 Knockout Raji Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population designed to disrupt the DNTTIP1 gene in a human B lymphocyte background. This product provides a loss-of-function model without selective clonal isolation, enabling pooled analysis of heterogeneous editing events across the cell population. The gene disruption is mediated by CRISPR/Cas9, targeting critical regions of DNTTIP1 to impair its expression and function. Researchers can utilize these cells to interrogate the pleiotropic roles of DNTTIP1 in DNA repair, V(D)J recombination, and chromatin dynamics. The polyclonal format retains genetic diversity, offering a robust system for studying gene function in a biologically relevant B-cell context.
The host Raji cell line is derived from a Burkitt’s lymphoma patient and represents an Epstein-Barr virus (EBV)-positive B lymphoblastoid model. These cells endogenously express B-cell lineage markers and recapitulate key aspects of lymphocyte biology, including immunoglobulin production and antigen presentation. Raji cells are extensively used in immunology and oncology research due to their rapid proliferation and susceptibility to DNA-damaging agents. Their transformed phenotype makes them particularly suitable for investigating genomic instability mechanisms and B-cell malignancy pathways, while retaining intact DNA repair machinery components such as DNA-PKcs and Ku70/Ku80, which are essential for non-homologous end joining (NHEJ).
DNTTIP1 encodes a terminal deoxynucleotidyltransferase (TdT)-interacting protein that orchestrates multiple nuclear processes. It physically binds TdT, modulating V(D)J recombination by influencing NHEJ complex assembly. DNTTIP1 also associates with MIDEAS and histone deacetylases HDAC1/HDAC2 within a chromatin-remodeling complex, linking DNA repair to epigenetic regulation and cell cycle progression. Upstream, transcription factors E2A, EBF1, and PAX5 regulate DNTTIP1 expression during lymphocyte development, while DNA damage sensors ATM and ATR signal to activate DNTTIP1-dependent repair pathways. Downstream consequences of DNTTIP1 action include altered activity of TdT, recruitment of DNA-PKcs and Ku70/Ku80 to DNA lesions, and modulation of cell cycle regulators such as p21 and cyclins. Through its interactions with NuRD subunits CHD4 and MTA1/2, DNTTIP1 further coordinates chromatin accessibility and transcriptional programs critical for genomic stability.
In the Raji B-cell context, loss of DNTTIP1 disrupts these multifaceted networks, providing a powerful tool to dissect mechanisms underlying B-cell transformation and therapy resistance. Since DNTTIP1 regulates NHEJ-mediated repair and cell cycle checkpoints, this knockout model can reveal vulnerabilities in lymphoma cells exposed to genotoxic chemotherapeutics or targeted inhibitors. Aberrant V(D)J recombination activity, although less prominent in mature B cells, may still contribute to genomic rearrangements in lymphomagenesis, making DNTTIP1 disruption relevant for studying aberrant repair events. Moreover, the interplay with HDAC1/2 and MIDEAS suggests roles in epigenetic dysregulation, a hallmark of many leukemias and lymphomas. Researchers can employ these cells to evaluate how DNTTIP1 deficiency affects proliferation, apoptosis, and response to DNA-damaging agents, thereby identifying potential synthetic lethal interactions or drug sensitizers.
Typical research applications include functional studies of V(D)J recombination using reporter assays, investigation of NHEJ efficiency through DNA damage response assays, and analysis of B-cell lymphoma pathogenesis via RNA-seq and ChIP-qPCR. The cells are validated for loss of DNTTIP1 protein expression by Western blotting, and downstream effects can be monitored by flow cytometry for cell cycle and apoptosis, RT-qPCR for gene expression changes, and co-immunoprecipitation to assess disrupted protein interactions. Drug sensitivity profiling with MTT assays can uncover altered chemosensitivity, while immunofluorescence enables visualization of repair protein localization. This model is also suited for genome stability research and immunodeficiency pathway exploration. For additional information and technical support, please contact Ascent Research.
