The GPR157 Knockout HEK293T Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout population targeting the human GPR157 gene in HEK293T cells. This loss-of-function model enables systematic investigation of the orphan G protein-coupled receptor GPR157, which has emerging roles in ciliary signaling and developmental pathways. The polyclonal format minimizes clonal artifacts and is well-suited for pooled functional screens and bulk biochemical assays, providing a robust and reproducible alternative to transient gene silencing approaches.
HEK293T is a human embryonic kidney epithelial cell line transformed with the SV40 large T antigen and constitutively expresses adenoviral E1A and E1B genes. These features confer high transfection efficiency and strong recombinant protein expression capacity, making HEK293T a staple platform for viral vector production, protein overexpression, and functional genomics. The cell line??s epithelial character and well-mapped signaling networks, including Hedgehog and cAMP pathways, render it an excellent host for interrogating gene function via CRISPR-mediated knockout.
GPR157 localizes to the primary cilium and functions as a negative regulator of the Hedgehog signaling pathway. Mechanistic studies suggest that GPR157 modulates intracellular cAMP levels, thereby influencing protein kinase A (PKA) activity and the proteolytic processing of Gli transcription factors. The receptor operates downstream of the Hedgehog receptor Patched (PTCH1) and upstream of the signal transducer Smoothened (SMO) and the Gli effectors. Known interacting partners include ??-arrestin2, which mediates receptor desensitization, and adenylyl cyclase, key for cAMP synthesis. Disruption of GPR157 leads to dysregulation of this ciliary signaling axis, affecting proper Hedgehog pathway output and potentially altering cAMP-dependent cellular processes.
In the context of HEK293T cells, which retain primary cilia and functional Hedgehog signaling components, GPR157 knockout creates a physiologically relevant model to study ciliary GPCR biology. The model permits detailed structure-function analyses through mutant rescue experiments and pharmacological manipulation of cAMP and Hedgehog pathways. Researchers can precisely dissect how GPR157 influences Gli transcriptional activity, ciliary morphology, and downstream gene expression programs. This system thus serves as a versatile platform for probing the molecular underpinnings of ciliopathy and cancer-related signaling.
The GPR157 Knockout HEK293T Polyclonal Cells are amenable to a broad range of experimental techniques. Western blotting and RT-qPCR allow confirmation of knockout and quantification of pathway component expression, while immunofluorescence enables visualization of ciliary protein localization. Functional assays include cAMP quantification, Gli-luciferase reporter assays for Hedgehog activity, and flow cytometry to analyze apoptosis and cell cycle changes. Migration assays further extend the phenotypic characterization. For further technical information, please contact Ascent Research.
