The GPRC5B Knockout HEK293T Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout population of HEK293T cells with targeted disruption of the GPRC5B gene. This loss-of-function model provides a heterogeneous cell pool for studying the orphan GPCR GPRC5B without requiring clonal isolation. The polyclonal format enables rapid experimental deployment in diverse assay systems while capturing population-level knockout effects relevant to signaling studies.
HEK293T cells are human embryonic kidney epithelial cells expressing SV40 large T antigen, ensuring high transfection efficiency and robust episomal plasmid replication. This cell line is a mainstay for recombinant protein production, lentivirus packaging, and transient transfection assays. The kidney epithelial origin also offers a relevant context for studies of renal signaling and cancer biology.
GPRC5B is a retinoic acid-inducible orphan GPCR that localizes to lysosomes, where it interacts with the Ragulator complex (LAMTOR1?C5) and Rag GTPases to regulate mTORC1 recruitment and activation. Induced by all-trans retinoic acid (ATRA) via RAR/RXR and regulated by SP1 and DNA methylation, GPRC5B couples nutrient sensing to downstream mTORC1 targets including S6K, 4E-BP1, and AKT, and also modulates MAPK/ERK and cAMP-PKA pathways through ??-arrestin and heterotrimeric G proteins. Its knockout disconnects retinoic acid signals from lysosomal nutrient sensing, impacting cell growth, autophagy, and proliferation.
In HEK293T cells, GPRC5B disruption impairs mTORC1-dependent phosphorylation cascades, altering protein synthesis and cell cycle progression. This provides a clean background for analyzing ATRA-dependent but GPRC5B-independent effects, as well as for evaluating how mTORC1 status influences recombinant protein yield and lentiviral production. The knockout cells thus serve as a platform for dissecting the crosstalk between retinoic acid signaling and growth pathways.
These cells are suited for Western blotting of mTORC1 markers (p-S6K, p-4E-BP1), immunofluorescence-based lysosomal localization studies, flow cytometric cell cycle analysis, RT-qPCR, and co-immunoprecipitation with Ragulator components. They also support autophagy assays (LC3 puncta) and proliferation measurements (MTT/BrdU). Applications span cancer biology, neurobiology, and drug screening targeting mTORC1 or GPCR modules. For more information, please contact Ascent Research.
