The GNAQ Knockout HeLa Polyclonal Cells provide a CRISPR/Cas9-edited polyclonal knockout cell population derived from the HeLa human cervical adenocarcinoma cell line. The cell pool contains a heterogeneous mixture of cells carrying loss-of-function mutations in the GNAQ gene, introduced by CRISPR/Cas9-mediated gene disruption. This polyclonal format allows robust, unbiased analysis of Gq signaling without clonal artifacts, making it suitable for functional genomics, pathway dissection, and compound screening targeting G??q-dependent processes.
HeLa cells, isolated from a cervical adenocarcinoma of patient Henrietta Lacks in 1951, are an immortalized epithelial line widely used in cancer research. They harbor integrated HPV18 DNA, resulting in E6- and E7-mediated inactivation of p53 and Rb, respectively. HeLa cells are highly proliferative, easy to transfect, and express a broad repertoire of signaling components, offering a versatile platform for CRISPR-edited knockout models. Their epithelial origin and transformed state provide a relevant background for studying GPCR signaling and tumor biology.
GNAQ encodes G??q, the alpha subunit of the heterotrimeric G protein Gq, which couples GPCR activation to phospholipase C beta (PLC??) stimulation. Upon receptor-mediated nucleotide exchange, G??q dissociates from G?¦? and activates PLC??, leading to IP3-mediated calcium release and DAG-dependent PKC activation. This cascade regulates MAPK/ERK, PI3K-AKT, and RhoA pathways, controlling cell growth, migration, and survival. G??q signaling is tightly modulated by RGS proteins and scaffolds such as GRK2, ??-arrestin, and caveolin-1. Activating mutations in GNAQ, commonly Q209L, drive constitutive signaling in uveal melanoma and other malignancies.
In HeLa cells, GNAQ knockout disrupts Gq-mediated responses downstream of endogenous GPCRs like endothelin receptors, M1/M3 muscarinic receptors, and 5-HT2 serotonin receptors, providing a clean background to study receptor-specific signaling. Loss of G??q abolishes calcium mobilization and PKC activation, enabling detailed assessment of Gq-dependent contributions to HeLa cell proliferation, migration, and oncogenic potential. This model is particularly valuable for exploring crosstalk between Gq signaling and HPV-driven transformation, as well as for dissecting Gq??s role in cervical adenocarcinoma biology.
This knockout cell pool supports a range of experimental applications, including Fluo-4 calcium flux assays to verify impaired GPCR signaling, IP3 accumulation measurements, and western blot analysis of phospho-ERK and phospho-PKC. Functional assays such as MTT cell proliferation and wound healing migration assays can assess phenotypic consequences of GNAQ loss. The cells also facilitate drug screening with Gq inhibitors like FR900359 and studies of GNAQ mutation-related oncogenesis. For additional information, please contact Ascent Research.
