DRD2 Knockout HAP1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population designed for loss-of-function studies of the dopamine receptor D2 gene. This product comprises a heterogeneous pool of edited cells harboring gene disruptions at the DRD2 locus, enabling robust functional investigation of DRD2-dependent signaling pathways without the limitations of single-cell clonal variability. Researchers can utilize this knockout model to dissect DRD2-mediated molecular mechanisms in a human genetic background.
The HAP1 cell line is a near-haploid human cell line derived from the KBM-7 chronic myeloid leukemia lineage, exhibiting adherent fibroblast-like morphology and a stable haploid karyotype. This genetic simplicity makes HAP1 cells particularly suited for genetic perturbation studies, as the presence of a single allele simplifies phenotypic interpretation and enhances knockout efficiency. As a model system, HAP1 cells support a wide range of biochemical and cell-based assays, including high-throughput screening, protein interaction analysis, and signal transduction studies, providing a versatile platform for investigating DRD2 function.
DRD2 encodes dopamine receptor D2, a G??i/o-coupled GPCR that inhibits adenylyl cyclase, reducing cAMP and PKA signaling upon dopamine binding. This decreases phosphorylation of DARPP-32 and modulates PP2A activity. Beta-arrestin-2 recruitment activates MAPK/ERK1/2 and AKT/GSK3?? pathways, influencing gene expression and neuronal plasticity. The receptor heterodimerizes with adenosine A2A receptors and interacts with GRK2, GRK3, calmodulin, and spinophilin, integrating dopaminergic and adenosine signaling. Downstream, DRD2 modulates Ca2+ and K+ channels and inhibits prolactin secretion.
In HAP1 cells, the loss of DRD2 expression creates a simplified experimental chassis to analyze dopamine receptor signaling independently of neuronal complexity. The polyclonal knockout population allows assessment of the average functional impact of DRD2 disruption across diverse mutations, mitigating clonal artifacts associated with monoclonal lines. This model is valuable for studying antipsychotic drug mechanisms, as DRD2 is the primary target of haloperidol, and the haploid background facilitates saturation mutagenesis screens and drug?Cgene interaction profiling to identify modulators of DRD2-dependent pathways.
Typical applications include cAMP accumulation assays, phospho-ERK1/2 western blotting, BRET assays for beta-arrestin translocation, radioligand binding, high-throughput drug screening, co-immunoprecipitation, and cell viability assays with antipsychotics. This model serves neuropsychiatric disease research, addiction biology, and dopaminergic signaling studies. For further information, please contact Ascent Research.
