The HRH1 Knockout HCT 116 Polyclonal Cells comprise a CRISPR/Cas9-edited polyclonal knockout population targeting the HRH1 gene in HCT 116 cells. This product delivers a heterogeneous pool of cells with disrupted HRH1 expression, providing a loss-of-function model without clonal selection. The polyclonal format captures the diversity of editing outcomes, making it suitable for experiments where population-level effects are desired.
HCT 116 is a human colorectal carcinoma cell line with epithelial morphology, derived from a male patient. These cells are near-diploid, display microsatellite instability, and harbor a KRAS G13D mutation. HCT 116 serves as an established system for colorectal cancer research, recapitulating key features such as dysregulated proliferation and signaling. The genetic background enables studies of oncogenic pathways and their interaction with GPCR signaling.
The HRH1 receptor, activated by histamine, couples to Gq/11 proteins to stimulate phospholipase C, generating IP3 and DAG. IP3 mobilizes intracellular calcium, while DAG activates PKC, which then phosphorylates ERK1/2. These kinases regulate transcription factors AP-1 and NF-??B, promoting expression of inflammatory and proliferative genes. HRH1 signaling is modulated by beta-arrestin recruitment and interacts with GRK2 and CaM kinase. Downstream targets include c-Fos and components of the MAPK pathway. Knockout of HRH1 abrogates this axis, enabling analysis of histamine-dependent cellular responses.
In HCT 116 cells, HRH1 disruption allows dissection of histamine??s role in colorectal cancer biology. The KRAS mutation and MSI status may intersect with HRH1-driven pathways, potentially influencing calcium mobilization, MAPK activation, and NF-??B-mediated inflammation. This model is valuable for investigating how histamine signaling contributes to tumor progression and inflammatory conditions like colitis. It also provides a platform to study receptor crosstalk and identify HRH1-specific therapeutic vulnerabilities.
Applications include quantitative RT-qPCR and western blotting to confirm HRH1 ablation and assess downstream signaling molecules. Functional assays such as calcium flux measurement, phospho-ERK ELISA, and NF-??B reporter assays can be performed following histamine stimulation. Cell proliferation and migration assays reveal HRH1-dependent effects on growth and motility. Antagonist treatment with H1 antihistamines validates receptor-specific pharmacology. These cells are ideal for GPCR signaling studies, inflammation research, and drug sensitivity screening in colorectal cancer. For further technical details, contact Ascent Research.
