The DOK1 Knockout HEK293T Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population in which the DOK1 gene has been disrupted to create a loss-of-function model. This product provides a heterogeneous pool of HEK293T cells carrying diverse DOK1 edits, enabling functional studies of DOK1 deficiency without clonal selection artifacts. The polyclonal format preserves population-level responses and is well-suited for pooled screens, signaling assays, and comparative analyses with parental controls.
The host cell line, HEK293T, is a widely used human embryonic kidney epithelial line that constitutively expresses the SV40 large T antigen. This expression facilitates high-level plasmid amplification and efficient viral production, making HEK293T cells a standard platform for protein expression, lentivirus and retrovirus packaging, and transient transfection-based assays. The cells’ epithelial origin and robust growth characteristics also support reproducible signaling experiments across a broad range of stimuli and pathway perturbations.
DOK1 encodes an adaptor protein that functions as a negative regulator of Ras-MAPK signaling and cell proliferation. Upon activation of upstream receptors such as EGFR, PDGFR, TCR, BCR, and the IL-2 receptor, DOK1 is phosphorylated by Src family kinases or JAK, creating docking sites for interacting partners. Through its interactions with RasGAP (RASA1), SHIP1, and Csk, DOK1 attenuates Ras-ERK1/2 and PI3K-AKT signaling, thereby modulating immune cell activation, cytokine responses, and tumor suppression. In T cells, DOK1 engagement downstream of the TCR helps set activation thresholds by limiting ERK and JNK phosphorylation.
In the HEK293T epithelial context, loss of DOK1 is expected to relieve negative feedback on Ras-MAPK and PI3K-AKT pathway activity, leading to enhanced basal and ligand-induced ERK1/2 phosphorylation. Because HEK293T cells do not require DOK1 for viability, the knockout model allows clean interrogation of DOK1-dependent signaling modules without confounding effects from endogenous hematopoietic-specific pathways. The polyclonal population is particularly advantageous for phenotypic screens and for dissecting DOK1’s contribution to NF-??B and JNK activation downstream of various receptors.
Researchers can employ this DOK1 knockout model in a variety of experimental contexts, including investigation of Ras-MAPK regulation, kinase inhibitor profiling, and tumor suppressor mechanism studies. Typical readouts include Western blotting for DOK1 and phospho-ERK, phospho-kinase arrays, RT-qPCR, cell proliferation assays, luciferase reporter assays, and co-immunoprecipitation of signaling complexes. The cells also serve as a valuable tool for functional genomics and for validating small-molecule inhibitors targeting upstream kinases. For further information, please contact Ascent Research.
