GPR107 Knockout HEK293T Polyclonal Cells provide a CRISPR/Cas9-mediated gene disruption of the orphan GPCR GPR107. This polyclonal knockout cell population is derived from HEK293T cells and offers a heterogeneous pool of edited cells for functional studies. The product serves as a loss-of-function model to dissect the roles of GPR107 in signal transduction and intracellular trafficking without requiring single-cell clonal isolation.
HEK293T is a human embryonic kidney epithelial cell line stably expressing the SV40 large T antigen. This transformation yields high transfection efficiency and robust protein production, making these adherent cells with epithelial morphology a preferred system for transient gene expression, virus packaging, and receptor pharmacology. Their well-characterized endogenous signaling machinery provides an optimal background for studying GPCR-mediated pathways.
GPR107 is predicted to be a seven-transmembrane domain orphan receptor. Although its ligand remains unknown, GPR107 is regulated by GPCR kinases (GRK2 and GRK5) and ??-arrestins (??-arrestin1/2). Downstream, it couples to heterotrimeric G proteins, including G??s, G??i, and G??q family members, modulating adenylate cyclase and phospholipase C to produce second messengers cAMP and IP3. These activate effectors such as protein kinase A (PKA), protein kinase C (PKC), and the MAPK/ERK cascade. Additionally, GPR107 interacts with retromer complex components VPS35, VPS26, and SNX1, as well as Rab GTPases Rab7 and Rab9, linking it to endosome-to-Golgi retrograde transport.
In the HEK293T background, endogenous GPCR expression and trafficking machinery provide a physiologically relevant context for probing GPR107 function. The polyclonal knockout population enables population-level analyses of gene disruption effects, avoiding clonal artifacts and facilitating robust statistical comparisons in cell-based assays. This model is particularly suited for examining how loss of a receptor that bridges signaling and endosomal sorting impacts cellular homeostasis in an epithelial setting.
Typical applications include cAMP and calcium mobilization assays to quantify second messenger changes, co-immunoprecipitation to assess interactions with ??-arrestins or retromer proteins, and immunofluorescence to visualize altered subcellular localization of trafficking components. Antibody internalization experiments reveal endocytic defects, while RNA-seq and RT-qPCR profile transcriptional consequences. Flow cytometry and western blotting provide complementary expression validation. These tools support orphan receptor deorphanization, signaling pathway dissection, and drug target validation. For further information, contact Ascent Research.
