The DOK5 Knockout HAP1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout population for loss-of-function studies of the DOK5 gene. This genetically heterogeneous pool of HAP1 cells carries targeted disruption of the DOK5 locus, enabling functional genomic analyses without clonal selection. The polyclonal format preserves population diversity while eliminating DOK5-dependent signaling, suited for pooled screening and pathway interrogation.
The HAP1 host cell line is a near-haploid human cell model derived from the KBM-7 chronic myelogenous leukemia line. These male, adherent fibroblast-like cells contain a single copy of most chromosomes, simplifying genome engineering and phenotype interpretation. Widely used in functional genomics, HAP1 cells provide a robust, manipulable platform for dissecting gene function in signal transduction.
DOK5 is an adaptor protein and positive regulator of neurotrophic and insulin signaling. Phosphorylated by receptor tyrosine kinases RET (GDNF) and TrkB (BDNF), DOK5 recruits SH2 domain-containing proteins CRK and NCK, propagating signals through the MAPK/ERK cascade via TrkB, SHC, GRB2, SOS, RAS, RAF, MEK, and ERK, promoting neurite outgrowth. In insulin signaling, DOK5 interacts with IRS1, modulating PI3K/AKT pathways. Src kinases also phosphorylate DOK5, integrating upstream inputs. Thus, DOK5 coordinates neurotrophic and metabolic signaling.
Disruption of DOK5 in HAP1 cells creates a clean system to study neurotrophin and insulin signaling without compensatory diploid mechanisms. The near-haploid nature ensures single genetic alterations are phenotypically unmasked, enabling clear dissection of DOK5’s role in MAPK/ERK activation and neurite outgrowth. This model is valuable for investigating TrkB and RET downstream events and insulin responsiveness. The adherent fibroblast-like morphology supports imaging-based assays for cytoskeletal dynamics.
This DOK5 knockout polyclonal population supports functional genomic screens, drug target validation, and mechanistic studies of neurotrophic signaling. Assays such as Western blotting for phospho-ERK, RT-qPCR, immunofluorescence for neurite dynamics, and co-immunoprecipitation of DOK5 interactors are applicable. Cells can be used in neurite outgrowth or insulin signaling activation assays. Researchers studying type 2 diabetes, neurodevelopmental disorders, or RET pathways will find this model powerful. For further details, contact Ascent Research.
