The HRAS Knockout 769-P Polyclonal Cells constitute a CRISPR/Cas9-edited polyclonal knockout cell population derived from the 769-P clear cell renal cell carcinoma line. These polyclonal knockout cells carry targeted disruption of the HRAS gene, abolishing expression of the HRAS small GTPase. The polyclonal format provides a heterogeneous pool of knockout alleles, offering a robust loss-of-function model for population-level studies without clonal selection bias, and is readily expandable for downstream applications.
The parental 769-P cell line is an established in vitro model of clear cell renal cell carcinoma (ccRCC), a common and aggressive kidney cancer subtype. 769-P cells retain hallmark features of ccRCC, including dysregulated hypoxia signaling and activation of oncogenic pathways. This cell line is widely employed to investigate ccRCC biology, therapeutic vulnerabilities, and mechanisms of tumor progression.
HRAS encodes a membrane-associated small GTPase that orchestrates signal transduction from cell surface receptors to intracellular effector cascades. In its active GTP-bound state, HRAS directly engages RAF1, PI3K, and RALGDS, thereby driving the MAPK/ERK, PI3K/AKT, and RALGDS signaling modules. Activation of HRAS occurs downstream of receptor tyrosine kinases such as EGFR and FGFR, facilitated by adaptor proteins like GRB2 and guanine nucleotide exchange factors including SOS1. HRAS is negatively regulated by GTPase-activating proteins such as NF1 and p120GAP, which accelerate GTP hydrolysis. The MAPK/ERK cascade, in which HRAS relays signals via RAF1, MEK1/2, and ERK1/2, controls cell proliferation and differentiation. Concurrently, the PI3K/AKT axis promotes survival and metabolic reprogramming, while RALGDS signaling contributes to cytoskeletal remodeling and migration.
In the context of 769-P ccRCC cells, HRAS knockout disrupts these convergent oncogenic pathways, providing a powerful tool to dissect RAS-dependent growth and survival mechanisms. Polyclonal knockout in this background is especially relevant because HRAS mutations or overexpression have been implicated in a subset of kidney tumors. By eliminating HRAS function, this model enables investigation of cancer cell addiction to RAS signaling and assessment of residual pathway activation through alternate RAS isoforms or bypass mechanisms. It also allows study of the interplay between HRAS loss and the tumor microenvironment, as well as the impact on other signaling nodes such as PI3K/AKT.
This polyclonal knockout cell population is suitable for diverse functional genomics and translational cancer research applications, including oncogene addiction studies, drug target validation, and synthetic lethality screening. Typical assays include phospho-ERK immunoblotting to monitor MAPK pathway activity, proliferation and colony formation assays to evaluate growth dependence, and migration assays to assess metastatic potential. Additionally, these cells can be used in RT-qPCR and western blotting to confirm gene disruption and downstream effector modulation. The HRAS Knockout 769-P Polyclonal Cells offer a rigorous tool for dissecting RAS signaling in ccRCC. For further technical specifications or ordering information, please contact Ascent Research.
