The DOK3 Knockout HAP1 Polyclonal Cells provide a CRISPR/Cas9-edited polyclonal knockout population designed for loss-of-function studies of the adaptor protein DOK3. This product comprises a heterogeneous pool of HAP1 cells harboring targeted disruptions in the DOK3 gene, enabling robust investigation of DOK3-dependent signaling without the need for clonal isolation. The polyclonal format is well-suited for high-throughput screening and pooled functional assays where population-level effects are informative.
HAP1 is a near-haploid, fibroblast-like adherent cell line derived from the KBM-7 chronic myeloid leukemia line. Its near-haploid karyotype simplifies gene editing and reduces the complexity of diploid compensation, making it an attractive host for knockout model generation. HAP1 cells maintain stable growth and are amenable to a range of standard cell biology techniques, including transfection, immunoblotting, and fluorescence imaging. These characteristics render them a preferred chassis for dissecting signaling networks where gene dosage effects are minimal.
DOK3 functions as a cytoplasmic adaptor and negative regulator of immunoreceptor signaling, particularly in B cells and macrophages. Following B-cell receptor (BCR) or Fc receptor engagement, DOK3 is phosphorylated by Src-family kinases such as Lyn and Syk. Once phosphorylated, DOK3 recruits the inositol phosphatase SHIP1 and the adaptor Grb2, which together attenuate downstream MAPK/ERK and NF-??B cascades. This scaffolding activity dampens calcium flux, proliferation, and transcriptional responses, thereby fine-tuning lymphocyte activation. Consistent with its inhibitory role, DOK3 signaling intersects with key nodes including Ras, BTK, PLC??2, and Csk.
In the HAP1 polyclonal knockout model, disruption of DOK3 provides a simplified cellular context to dissect the molecular requirements for negative feedback in signaling. Although HAP1 is not of lymphoid origin, its genetic plasticity enables reconstitution of immunoreceptor pathways via ectopic expression of relevant receptors and kinases. Researchers can introduce BCR components or stimulate endogenous growth factor receptors to probe DOK3-dependent modulation of ERK and NF-??B. This model thus serves as a versatile platform for structure?Cfunction analyses of DOK3 and screening of adaptor-mediated signal termination.
Typical applications include phospho-ERK flow cytometry to assess signaling strength upon receptor stimulation, co-immunoprecipitation to map DOK3?CSHIP1 or DOK3?CGrb2 interactions, and NF-??B reporter assays to measure transcriptional outcomes. The polyclonal population is also suitable for proliferation assays and calcium flux measurements following BCR-like stimulation. Furthermore, the knockout cells support drug target validation studies in autoimmune and inflammatory disease contexts, where DOK3??s regulatory role makes it a candidate for therapeutic intervention. For further experimental guidance or customization options, please contact Ascent Research.
