The DTWD2 Knockout Jurkat Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from Jurkat T lymphocytes. This product features targeted disruption of the DTWD2 gene, which encodes a substrate recognition subunit of the CUL4-DDB1-RBX1 E3 ubiquitin ligase complex. The polyclonal format provides a genetically diverse pool of knockout cells, enabling robust functional analyses without clonal selection. This model serves as a powerful tool for dissecting DTWD2-dependent ubiquitination and protein degradation pathways.
The parental Jurkat cell line is a suspension lymphoblast line originally established from the peripheral blood of a 14-year-old male with acute T-cell leukemia. Jurkat cells are a canonical model for T-cell receptor signaling, apoptosis, and leukemia biology, with extensive characterization of their proteome and signaling networks. Their lymphoid origin and well-defined apoptotic machinery make them an optimal host for investigating ubiquitin ligase components that govern protein stability and cellular fate in T cells.
DTWD2 functions as a substrate receptor that recruits target proteins to the CUL4-DDB1-E3 ligase complex. It interacts directly with DDB1 and indirectly with the scaffold cullin proteins CUL4A or CUL4B and the RING domain protein RBX1, which activates ubiquitin conjugation from E2 enzymes to substrates. This ubiquitination tags proteins for recognition and degradation by the 26S proteasome, constituting a key regulatory node in the ubiquitin-proteasome system. The identity of specific downstream targets remains largely unknown, and the upstream regulators controlling DTWD2 expression are not yet defined. Representative molecular components involved in this pathway include CUL4A, DDB1, RBX1, DTWD2, ubiquitin, and the 26S proteasome.
In Jurkat cells, protein degradation via the ubiquitin-proteasome system is essential for modulating T-cell activation, proliferation, and apoptosis. Perturbations in E3 ligase function can dysregulate these processes and contribute to leukemogenesis. DTWD2 knockout in this leukemia-relevant background allows researchers to examine how loss of this substrate receptor alters protein turnover, signal transduction, and apoptotic sensitivity. This model is particularly valuable for exploring connections between CUL4-DDB1-mediated ubiquitination and T-cell acute lymphoblastic leukemia pathology, as well as for evaluating the impact on cell cycle progression and stress responses.
DTWD2 Knockout Jurkat Polyclonal Cells are suited for a wide range of functional studies, including identification of CUL4-DDB1 substrates, protein turnover analysis, and CRISPR-based functional genomics screens. They can be employed in proteomic profiling to map DTWD2-dependent ubiquitination events and in drug sensitivity assays to assess chemotherapeutic responses in a leukemia context. Representative assays include Western blotting for ubiquitin conjugates, RT-qPCR for transcript validation, co-immunoprecipitation of ligase complexes, in vitro ubiquitination reactions, flow cytometry to measure apoptosis and cell cycle parameters, and proteasome inhibition experiments. For further information, please contact Ascent Research.
