The DTX3 Knockout HAP1 Polyclonal Cells comprise a CRISPR/Cas9-edited polyclonal knockout cell pool designed to disrupt the DTX3 gene in the near-haploid HAP1 cell line. Unlike clonal isolates, this polyclonal population includes a spectrum of genetic variants, offering a comprehensive platform for functional analysis of DTX3-dependent pathways without the confounding effects of clonal selection.
HAP1 cells, originally derived from the KBM-7 chronic myeloid leukemia line, harbor the BCR-ABL oncogenic fusion and possess a near-haploid karyotype. This genetic simplicity enables efficient CRISPR/Cas9-mediated knockout and makes HAP1 a widely adopted system for haploid genetic screens and cancer signaling research.
DTX3 is an E3 ubiquitin ligase that critically modulates Wnt/??-catenin and Notch signaling. It directly ubiquitinates ??-catenin, targeting it for proteasomal degradation and thereby suppressing Wnt-driven transcription. DTX3 also interacts with Notch receptors and other Deltex family members (DTX1, DTX2, DTX4) and is regulated by upstream signals such as Notch ligands (JAG1, DLL1), NICD, and Wnt ligands (Wnt3a). DTX3 knockout consequently stabilizes ??-catenin, leading to increased Wnt pathway activity and potential crosstalk with Notch signaling.
In the HAP1 chronic myeloid leukemia background, DTX3 loss creates a model of constitutive Wnt activation relevant to leukemogenesis and solid tumors. The haploid nature of these cells facilitates synthetic lethality screening and genome-wide knockout studies to uncover genetic dependencies linked to ??-catenin accumulation, thereby aiding identification of therapeutic targets.
These polyclonal knockout cells are suitable for a variety of assays, including TOP/FOP flash luciferase reporter assays for Wnt activity, Western blotting for DTX3 and ??-catenin, RT-qPCR of Wnt target genes (e.g., AXIN2, MYC), and co-immunoprecipitation to probe protein interactions. Drug sensitivity profiling, cell proliferation assays, and haploid genetic screens further expand their utility in oncology research. For additional technical information, please contact Ascent Research.
