The GPRIN2 Knockout HeLa Polyclonal Cells are a CRISPR/Cas9-edited polyclonal cell population derived from the HeLa cervical adenocarcinoma line, engineered for disruption of the GPRIN2 gene. This pooled knockout model, containing a heterogeneous array of loss-of-function alleles, enables robust population-level analysis of GPRIN2-dependent signaling. The product is tailored for investigations into GPCR pathways, neurite outgrowth mechanisms, and cytoskeletal regulation.
HeLa cells, an HPV-18-positive cervical adenocarcinoma line, are immortalized via E6 and E7 oncoprotein-mediated inactivation of p53 and Rb. This highly proliferative, aneuploid cell model is a workhorse for cancer research and protein expression, offering high transfection efficiency and well-characterized signaling networks, including GPCR cascades. Its robust growth and tractability make it an ideal host for studying the functional consequences of GPRIN2 knockout.
GPRIN2 operates as a G protein-regulated inducer of neurite outgrowth downstream of Gi/o-coupled GPCRs. It modulates adenylyl cyclase activity and cAMP/PKA signaling, while also activating the MAPK/ERK pathway via phosphorylation of ERK1/2 and downstream transcription factor CREB. GPRIN2 interacts with 14-3-3 proteins, PKA, and cytoskeletal adaptors to reorganize MAP2 and Tau, driving neurite extension. It is activated by neurotrophins (BDNF, NGF) and intersects with Bcl-2 family proteins to link survival cues to morphological differentiation. Disruption of GPRIN2 in HeLa cells thus provides a clean background to dissect these molecular interactions and signaling branches.
Within HeLa cells, which lack a neuronal phenotype, GPRIN2 knockout serves as a valuable tool for parsing GPCR signaling modules related to cAMP and ERK without neuronal differentiation artifacts. These knockout cells are particularly suited for migration and invasion assays, as GPRIN2 may influence cytoskeletal dynamics relevant to cancer cell motility. In addition, HeLa cells can be induced to form neurite-like processes via overexpression of key factors, enabling neurite outgrowth studies in a genetically tractable system. The polyclonal nature ensures a broad representation of mutations, increasing the robustness of phenotypic analyses.
Typical applications include western blotting, RT-qPCR, and immunofluorescence for assessing gene loss and downstream effects; cAMP and phospho-ERK assays for pathway activity; and live-cell migration and invasion assays. The cells are compatible with co-immunoprecipitation, RNA-seq, and drug screening platforms for neuropsychiatric disorders. The heterogeneous knockout pool enhances statistical power, making it ideal for functional genomics and compound profiling. For additional information and validation data, please contact Ascent Research.
