The DOK2 Knockout HAP1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population for loss-of-function studies of DOK2 in a human near-haploid background. This product provides a genetically disrupted DOK2 model from the HAP1 cell line, enabling investigation of DOK2-dependent signaling networks and tumor-suppressive mechanisms. The polyclonal population contains diverse editing events while maintaining consistent gene disruption, suitable for functional genomic screens and mechanistic analyses without clonal isolation.
The HAP1 line is a near-haploid human cell line derived from the KBM-7 chronic myeloid leukemia (CML) line, retaining a near-haploid karyotype with disomy only for chromosome 8. This genetic simplicity facilitates manipulation and reduces confounding effects from heterozygosity. HAP1 cells express the BCR-ABL fusion oncogene, providing an endogenous tyrosine kinase signaling context relevant to leukemogenesis. Widely used for CRISPR-based functional genomics, high-throughput screening, and signaling studies, HAP1 offers robust growth and ease of genetic perturbation.
DOK2 is an adaptor protein that negatively regulates tyrosine kinase signaling. It is phosphorylated by upstream kinases including BCR-ABL, JAK2, and Src family members (Lyn, Fyn), creating docking sites for inhibitory effectors. DOK2 recruits RASA1 (RASGAP) to inactivate Ras and attenuate MAPK/ERK signaling, and interacts with INPP5D (SHIP) to downregulate PI3K/AKT. Additionally, DOK2 recruits Csk to suppress Src kinases, establishing a negative feedback loop that limits proliferative and survival signals from cytokine receptors (EPO, IL-3) and antigen receptors (TCR, BCR). Thus, DOK2 acts as a molecular brake on oncogenic and immune receptor pathways.
In HAP1 cells, DOK2 disruption removes a critical negative feedback mechanism that restrains BCR-ABL-driven signaling. This knockout model is ideal for dissecting the interplay between DOK2 and BCR-ABL, as these cells depend on constitutive kinase activity. Loss of DOK2 can lead to hyperactivation of Ras/MAPK and PI3K/AKT, increased proliferation, and altered sensitivity to tyrosine kinase inhibitors like imatinib. Therefore, these polyclonal knockout cells provide a physiologically relevant system to study tumor suppression in CML and adaptive resistance in kinase-dependent cancers.
The DOK2 Knockout HAP1 Polyclonal Cells support diverse experimental applications, including functional validation of DOK2 as a tumor suppressor, analysis of negative feedback in tyrosine kinase signaling, and investigation of Ras/MAPK and PI3K/AKT modulation. They are suited for drug resistance studies (e.g., imatinib sensitivity) and protein interaction mapping via co-immunoprecipitation of DOK2 with RASA1 or SHIP. Typical assays include Western blotting for phospho-ERK and phospho-AKT, RT-qPCR, phosphotyrosine profiling, cell proliferation and apoptosis assays, and flow cytometry for cell cycle analysis. For further information, please contact Ascent Research.
