The GRK6 Knockout HAP1 Polyclonal Cells provide a CRISPR/Cas9-mediated polyclonal knockout population targeting the GRK6 gene in near-haploid HAP1 cells. As a pool of cells with diverse GRK6-disrupting alleles generated by non-homologous end joining, this format allows bulk functional studies and pooled genetic screens without the influence of clonal selection. The polyclonal mixture yields a population-averaged loss-of-function phenotype, suitable for assessing signaling pathway alterations and receptor regulation in a genetically tractable background.
HAP1 is a near-haploid human cell line originating from KBM-7 chronic myeloid leukemia blast crisis cells. Its haploid karyotype simplifies gene disruption because only a single allele needs to be targeted to achieve functional knockout, circumventing diploid redundancy. HAP1 retains hematopoietic features, expresses endogenous GPCRs including chemokine receptors, and activates downstream kinases such as ERK1/2 and Akt. This makes it an apt model for investigating kinase-dependent pathways in a leukemic context, particularly those linked to cell migration and signal desensitization.
GRK6 belongs to the G protein-coupled receptor kinase family and preferentially phosphorylates activated GPCRs, notably CXCR4 and CCR5. Phosphorylation promotes beta-arrestin 2 recruitment, which sterically hinders G protein coupling and facilitates clathrin-AP-2-mediated endocytosis. GRK6 activity is modulated by GPCR ligands, protein kinase C, Src kinases, and TGF-beta, and it interacts with G?¦? subunits, PIP2, and calmodulin. Downstream, beta-arrestin scaffolds the ERK1/2 and Akt cascades, regulating transcriptional programs that control immune cell migration, chemotaxis, and survival??processes central to inflammation and cancer progression.
In the HAP1 background, disruption of GRK6 abrogates the primary mechanism for ligand-induced chemokine receptor desensitization and internalization. This enables precise evaluation of how GRK6 deficiency alters CXCR4- and CCR5-mediated signaling, calcium responses, and cell migration. Given the CML derivation, the model also permits exploration of GRK6??s role in leukemic cell trafficking and malignant hematopoiesis, bridging studies of normal immune cell function and oncogenic signaling.
This polyclonal knockout tool is applicable to functional genomics screens, GPCR signaling analyses, and drug target validation in inflammation, autoimmunity, cancer, and cardiovascular disorders. Typical experimental readouts include Western blotting and RT-qPCR for GRK6 expression, chemotaxis and invasion assays, phospho-ERK flow cytometry, GPCR internalization assays, co-immunoprecipitation of beta-arrestin?CGPCR complexes, and NF-??B reporter assays. For additional product details or inquiries, please contact Ascent Research.
