The GPR156 Knockout HEK293T Polyclonal Cells constitute a CRISPR/Cas9-edited polyclonal knockout cell population targeting the GPR156 gene. This product comprises a heterogeneous pool of HEK293T cells with diverse gene disruptions introduced by non-homologous end joining, yielding a loss-of-function model for the orphan G protein-coupled receptor GPR156. The polyclonal format avoids clonal selection biases and provides a robust cellular system for interrogating receptor-mediated signaling pathways in a human embryonic kidney background.
HEK293T cells are derived from human embryonic kidney tissue and stably express the SV40 large T antigen, which permits episomal replication of plasmids containing the SV40 origin. This property, combined with high transfection efficiency, establishes HEK293T as a premier host for protein expression, viral packaging, and gene editing. The cells are widely employed in GPCR research because they provide a relatively silent endogenous receptor landscape, allowing clean interpretation of recombinant signaling events.
GPR156 is an orphan GPCR with no known endogenous ligands, yet it has been genetically linked to autosomal recessive deafness (DFNB) and vestibular dysfunction. Upon putative activation, GPR156 couples to heterotrimeric G proteins, including G?? q/11, G?? s, and G?? i/o, to regulate intracellular second messengers such as cAMP and Ca2+. Downstream effectors encompass phospholipase C, adenylyl cyclase, protein kinase A, protein kinase C, and MAP kinase cascades. The receptor also interacts with ??-arrestin 1/2 and G protein-coupled receptor kinases (GRKs), which modulate its desensitization and trafficking.
Knockout of GPR156 in HEK293T cells ablates its potential to influence these signaling networks, creating a defined model to dissect its cellular functions. The absence of confounding endogenous GPCRs in the host line ensures that observed phenotypes directly result from GPR156 disruption. This system is particularly valuable for studying how loss of the receptor affects second messenger dynamics, transcriptional programs, and cytoskeletal organization, thereby advancing understanding of the molecular pathology underlying hereditary hearing loss.
This knockout cell population is suitable for a range of applications, including functional characterization of orphan receptors, elucidation of auditory signaling mechanisms, high-throughput screening for GPR156 modulators, and validation of therapeutic targets for sensorineural deafness. Researchers typically employ western blotting, RT-qPCR, cAMP accumulation assays, intracellular calcium measurements, luciferase reporters, immunofluorescence, and flow cytometry to analyze the knockout phenotype. For further information or to discuss custom projects, contact Ascent Research.
