The GPRC5A Knockout HT29 Polyclonal Cells are a genetically diverse population of HT29 human colorectal adenocarcinoma cells generated by CRISPR/Cas9-mediated disruption of the GPRC5A gene. This polyclonal format eliminates the bias associated with single-cell cloning, offering a more representative loss-of-function model that faithfully reflects the heterogeneous nature of tumor cell populations. This product is designed for investigators seeking to dissect the role of GPRC5A in colorectal cancer signaling and to validate its tumor-suppressive functions.
The parental HT29 cell line, established from a 44-year-old female with colorectal adenocarcinoma, displays epithelial morphology and serves as a standard intestinal epithelial model for cancer research. It is extensively employed to investigate tumor proliferation, differentiation, apoptosis, drug resistance, and metastasis, and it can be induced to undergo enterocytic differentiation in response to specific stimuli, making it a versatile platform for anti-cancer drug screening.
GPRC5A is a retinoic acid-inducible orphan G protein-coupled receptor that functions as a tumor suppressor by negatively regulating NF-??B and STAT3 signaling. Mechanistically, it inhibits the IKK complex, thereby reducing phosphorylation of NF-??B p65 and transcription of downstream targets such as IL6, TNF, BCL2, and CCND1, while also attenuating STAT3-driven expression of MYC and VEGF. GPRC5A directly interacts with IKK??/??, NF-??B p65, STAT3, and the TGF-?? receptor/Smad2/3 complex, thereby integrating multiple oncogenic inputs. Its expression is transcriptionally activated by retinoic acid through RAR/RXR nuclear receptors, placing GPRC5A at a key node between nuclear receptor and inflammatory signaling.
In HT29 colorectal adenocarcinoma cells, genetic disruption of GPRC5A leads to constitutive activation of NF-??B and STAT3 signaling, resulting in enhanced cell proliferation, resistance to apoptotic stimuli, and increased invasive capacity??phenotypes that closely resemble advanced colorectal carcinomas. This knockout model thus provides a clinically relevant system for dissecting the molecular mechanisms driving colorectal cancer progression and for assessing the efficacy of therapeutic agents targeting these oncogenic cascades.
Researchers can employ this polyclonal knockout population in a comprehensive suite of functional assays, including Western blot, RT-qPCR, NF-??B luciferase reporter assays, phospho-p65/STAT3 analysis, MTT/CCK-8 proliferation assays, Annexin V apoptosis assays, and Transwell migration/invasion assays. The model is also highly suitable for high-throughput drug sensitivity screens and genome-wide transcriptomic profiling by RNA-seq. For additional product information or to discuss customization options, please contact Ascent Research.
