The GPER1 Knockout Ca Ski Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the Ca Ski human cervical carcinoma line. They provide a targeted loss-of-function model for GPER1 (G protein-coupled estrogen receptor 1) via CRISPR/Cas9-mediated gene disruption, generating a heterogeneous pool with ablated GPER1 expression. This polyclonal format enables cost-effective estrogen signaling studies in a cervical cancer background without single-cell cloning.
The host cell line, Ca Ski, is an HPV-16-positive cervical squamous cell carcinoma epithelial line derived from a metastatic site. It retains integrated HPV-16 genomes and expresses viral oncoproteins E6 and E7, serving as a key model for HPV-driven cervical cancer, metastasis, and estrogen-responsive signaling. Ca Ski cells express both ER?? and GPER1, responding to estrogens with proliferative and migratory responses, making them ideal for studying membrane and nuclear estrogen receptor interplay.
GPER1 is a G protein-coupled receptor that mediates rapid non-genomic estrogen signaling. Activation by 17??-estradiol or the agonist G-1 triggers coupling to G??s and G??i, and recruits ??-arrestin-2, leading to EGFR transactivation via SHC and GRB2. This initiates the RAS?CRAF?CMEK?CERK1/2 cascade, phosphorylating ERK1/2 and inducing c-Fos and cyclin D1. Concurrently, GPER1 stimulates G??s?CAC?CcAMP?CPKA, phosphorylating CREB, and activates PI3K/AKT signaling. Downstream effectors include MMP-9 and cyclin D1. The receptor is modulated by tamoxifen and ICI 182,780.
In Ca Ski cells, GPER1 drives estrogen-induced proliferation, migration, and survival. Its knockout abolishes rapid signaling, preventing EGFR transactivation and ERK1/2 and AKT activation. This model enables investigation of GPER1??s role in HPV-positive cervical cancer progression, including cooperation with viral oncogenes and metastatic behavior, and allows dissection of crosstalk with nuclear estrogen receptors.
Typical applications include elucidating non-genomic estrogen signaling, screening selective GPER1 modulators, investigating chemoresistance, and studying HPV oncogenesis. Common assays include western blotting for phospho-ERK1/2 and phospho-AKT, RT-qPCR for FOS and CCND1, Transwell migration/invasion, calcium imaging, cAMP ELISA, and drug sensitivity testing with G-1 and tamoxifen. For further information, contact Ascent Research.
