GNG4 Knockout HAP1 Polyclonal Cells constitute a CRISPR/Cas9-edited polyclonal knockout cell population targeting the GNG4 gene in the near-haploid HAP1 line. The product provides a heterogeneous mixture of GNG4-disrupted alleles for robust loss-of-function studies. The GNG4 gene encodes the gamma-4 subunit of heterotrimeric G proteins, a key component of G beta-gamma signaling in GPCR pathways.
HAP1 is an adherent, fibroblast-like cell line derived from KBM-7 chronic myeloid leukemia cells, characterized by a near-haploid karyotype that simplifies gene disruption. HAP1 expresses the BCR-ABL1 fusion oncogene, making it particularly relevant for CML research. Its haploid genome reduces the need for biallelic editing and minimizes confounding compensatory mutations, commonly used in functional genomics screens and signal transduction studies.
GNG4 pairs with G beta subunits (GNB1, GNB2, GNB3) to form a stable beta-gamma dimer. Upon GPCR activation by ligands such as dopamine (via DRD2) or adrenergic (via ADRB2) agonists, G alpha subunits (e.g., GNAI1, GNAO1) exchange GDP for GTP and dissociate, liberating the beta-gamma complex. GNG4-containing beta-gamma directly activates phospholipase C-beta (PLCB2, PLCB3), phosphoinositide 3-kinase (PIK3CA, PIK3CG), and GRK2. This triggers IP3-mediated calcium release, DAG/protein kinase C, and PI3K-AKT and RAS-RAF-MEK-ERK cascades, ultimately regulating cell proliferation, survival, and migration.
In HAP1, GNG4 knockout disrupts beta-gamma-dependent signaling to PLC-beta and PI3K, providing a clean background to assess the gamma-4 subunit??s specific contributions. The leukemic origin and BCR-ABL1 expression allow studies of GPCR pathway crosstalk with oncogenic kinase signaling. Reduced calcium flux, AKT phosphorylation, and ERK activation can be monitored to evaluate GNG4??s role in CML cell growth and apoptosis resistance, with the polyclonal format ensuring diverse representation of editing outcomes.
This cell population supports calcium flux assays, phospho-ERK/AKT ELISA, western blotting, and RT-qPCR to quantify GPCR signaling outputs. It is suitable for cell migration/invasion assays, co-immunoprecipitation for beta-gamma interactome mapping, and BRET/FRET-based biosensor monitoring of G protein activation. The cells integrate easily into high-throughput screening for drug target validation or synthetic lethal interactions in leukemia. For further details or technical support, contact Ascent Research.
