The HRH1 Knockout 786-O Polyclonal Cells product from Ascent Research provides a CRISPR/Cas9-edited polyclonal knockout cell population derived from the 786-O human clear cell renal cell carcinoma line, with targeted disruption of the HRH1 gene. This polyclonal mixture captures a range of loss-of-function mutations within the cell pool, reflecting the genetic heterogeneity often observed in tumor populations and eliminating the need for clonal isolation. The model is suitable for applications requiring a genetically diverse background while maintaining the key oncogenic features of the parental cell line.
The 786-O host cell line is a widely used model of human renal clear cell carcinoma, characterized by a biallelic mutation in the von Hippel-Lindau (VHL) tumor suppressor gene. This mutation results in constitutive stabilization of hypoxia-inducible factors (HIF-1?? and HIF-2??), creating a pseudo-hypoxic transcriptional state that drives angiogenesis, metabolic reprogramming, and tumor progression. The VHL-deficient background provides a highly relevant cellular context for investigating receptor-mediated signaling pathways implicated in kidney cancer.
HRH1 encodes the histamine H1 receptor, a Gq/11-coupled GPCR that mediates histamine-induced signaling. Upon histamine binding, the receptor activates G??q/11 proteins (GNAQ/GNA11), which stimulate phospholipase C?? (PLC??) to hydrolyze PIP2 into the second messengers IP3 and DAG. IP3 triggers calcium mobilization from intracellular stores, while DAG activates protein kinase C (PKC). The resulting PKC and calcium signals converge on the MAPK/ERK cascade (RAF/MEK/ERK) and promote NF-??B nuclear translocation, driving transcription of pro-inflammatory cytokines (IL-6, IL-8), COX-2, and c-Fos. Receptor desensitization is mediated by GRK2 phosphorylation and subsequent recruitment of ??-arrestins (ARRB1 and ARRB2). Transcriptional regulators AP-1 and NF-??B, themselves downstream effectors, also provide feedback control of receptor expression.
In the 786-O renal carcinoma context, disruption of HRH1 expression is expected to impair histamine-induced Gq/11-PLC?? signaling, attenuating IP3-mediated calcium release and DAG-dependent PKC activation. Consequently, downstream ERK1/2 phosphorylation and NF-??B transcriptional activity are diminished, likely reducing the expression of pro-inflammatory and pro-tumorigenic mediators. This model thus enables precise dissection of HRH1-dependent pathways that may influence tumor cell proliferation, migration, and the inflammatory tumor microenvironment.
Typical applications include studying histamine receptor signaling in renal cell carcinoma, investigating HRH1-mediated inflammatory and allergic pathways in a ccRCC background, and screening for H1 receptor antagonists. Compatible assays encompass western blotting and RT-qPCR for target and pathway analysis, calcium flux and NF-??B luciferase reporter assays for signaling readouts, phospho-ERK detection for MAPK activation, and functional assays such as cell viability, Transwell migration/invasion, and RNA-seq transcriptional profiling. For further technical details or to discuss custom applications, please contact Ascent Research.
