The GNG12 Knockout Jurkat Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from Jurkat human T lymphocytes, designed to disrupt the GNG12 gene. This heterogeneous population provides a loss-of-function model for studying GNG12-dependent signaling without clonal bias. The knockout eliminates expression of the gamma-12 subunit, a critical component of G?¦? dimers, enabling dissection of G protein signaling in T cell biology.
Jurkat cells are an immortalized human CD4+ T-lymphocyte line originating from acute T cell leukemia, widely used to investigate T cell activation, chemokine signaling, and HIV infection. Their well-characterized GPCR and MAPK pathways make them ideal for interrogating GNG12 function. As a suspension culture, these cells facilitate high-throughput genetic and pharmacological studies in immunology and cancer research.
GNG12 encodes the gamma-12 subunit that associates with G?? proteins (GNB1-4) to form G?¦? dimers, which anchor to the plasma membrane and partner with G?? subunits (GNAI, GNAQ, GNAS) to relay signals from GPCRs like CCR5 and CXCR4. Upon stimulation, dissociated G?¦? activates effectors such as PLC??, PI3K, and ion channels, propagating second messengers (IP3, DAG, calcium) and activating PKC. It also regulates adenylyl cyclase?CcAMP?CPKA and MAPK cascades (ERK1/2, JNK, p38), culminating in NF-kB and CREB transcriptional responses.
In Jurkat T cells, GNG12-dependent G?¦? signaling governs chemokine-induced migration, adhesion, proliferation, and cytokine production. Knocking out GNG12 uncouples G?¦? from its effectors, allowing dissection of subunit-specific contributions to T cell activation and leukemogenesis. This model is relevant to cancer, inflammation, and immunodeficiency, where aberrant GPCR and MAPK signaling drive disease. The Jurkat background enables correlation of GNG12 loss with altered phosphorylation of AKT and ERK, providing mechanistic insights.
Applications include Transwell migration assays for chemotaxis, calcium flux measurements, and SRE-luciferase reporter assays to monitor transcriptional output. The cells support western blotting for phospho-ERK, phospho-AKT, and phospho-JNK, flow cytometry for activation markers, co-immunoprecipitation of G protein subunits, and RT-qPCR for cytokines like IL-2 and TNF. These tools validate drug targets in G protein-driven leukemia and inflammation. For further details, contact Ascent Research.
