The GPR156 Knockout HeLa Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population designed for targeted disruption of the GPR156 gene in human HeLa cells. This loss-of-function model enables detailed investigation of the orphan class C G protein-coupled receptor GPR156, supporting studies into its still poorly understood signaling mechanisms. The polyclonal format preserves allelic heterogeneity, offering a population-based view of gene function that avoids clonal artifacts.
HeLa cells are an immortalized human epithelial cell line derived from a cervical adenocarcinoma, characterized by the presence of HPV18 DNA, inactivation of the tumor suppressor p53, and constitutive reactivation of telomerase. This extensively characterized cell line is a mainstay in cancer research due to its robust proliferation and ease of genetic manipulation, providing a relevant platform for examining tumor suppressors in a transformed context.
GPR156 is an orphan class C GPCR proposed to act as a tumor suppressor by coupling primarily to Gi/o proteins, leading to inhibition of adenylyl cyclase, reduced cAMP levels, and attenuation of PKA activity. This signaling converges on the MAPK/ERK cascade, with ERK1/2 phosphorylation as a key effector node. GPR156 may also engage ??-arrestin-2, facilitating scaffolding of additional signaling complexes or receptor internalization, and potentially heterodimerize with related class C GPCRs. Transcriptional regulation through CREB couples GPR156 activity to genes governing proliferation and migration.
In the HeLa background, where p53 function is compromised, GPR156 knockout is anticipated to further lift growth-suppressive constraints, thereby enhancing cell proliferation and migratory potential. This model allows dissection of GPR156-dependent signaling pathways that intersect with cancer-relevant processes, providing a controlled system to compare phospho-ERK levels and CREB activation between wild-type and edited populations.
Key applications include orphan GPCR functional studies, cancer cell biology investigations, and drug target identification campaigns. Users can evaluate functional consequences using MTT-based viability assays, Transwell migration and invasion assays, cAMP accumulation measurements, RT-qPCR for target gene expression, and Western blotting for phospho-ERK1/2. This polyclonal knockout population is well-suited for high-throughput screening to deorphanize GPR156 or identify modulators. For additional information or custom requests, please contact Ascent Research.
