The HRAS Knockout 143B Polyclonal Cells product consists of a heterogeneous population of CRISPR/Cas9-edited human osteosarcoma cells designed to disrupt the HRAS gene. This polyclonal knockout cell pool provides a biologically relevant loss-of-function model without clonal isolation, retaining genetic diversity that may better recapitulate tumor heterogeneity. The targeted gene disruption in the HRAS locus abrogates expression of the HRAS small GTPase, enabling robust interrogation of its signaling functions in a cancer-relevant background.
Derived from the highly tumorigenic 143B osteosarcoma cell line??an aggressive subclone of the HOS (TE85) lineage??these cells exhibit rapid proliferation and pronounced metastatic capability in vitro and in vivo. The 143B line is widely employed as a model for human bone tumorigenesis due to its faithful recapitulation of osteosarcoma growth, invasion, and metastatic dissemination. Its established use in xenograft studies and genetic manipulation makes it an ideal host for studying oncogene addiction and tumor suppressor mechanisms.
HRAS encodes a small GTPase that cycles between an inactive GDP-bound state and an active GTP-bound state, functioning as a molecular switch downstream of receptor tyrosine kinases such as EGFR and FGFR. Upon activation, HRAS-GTP engages multiple effector pathways, prominently including the MAPK/ERK cascade via direct interaction with RAF kinases (BRAF, CRAF) and the PI3K/AKT pathway through p110 catalytic subunits. Additionally, HRAS signals through RalGDS to activate Ral GTPases and phospholipase C?? to mobilize calcium and diacylglycerol. Adaptor proteins such as GRB2 and SOS facilitate HRAS nucleotide exchange, while GTPase-activating proteins like NF1 promote its inactivation.
Ablation of HRAS in 143B cells eliminates a critical node in oncogenic RAS signaling, disrupting downstream ERK1/2 and AKT phosphorylation and impairing transcriptional programs that drive uncontrolled proliferation, survival, and motility. This knockout model is particularly relevant for dissecting HRAS-dependent tumorigenic mechanisms in osteosarcoma, where mutations or overexpression of RAS pathway components are implicated. By uncoupling HRAS from its effectors, researchers can delineate signal rewiring, identify compensatory pathways, and evaluate therapeutic vulnerabilities specific to RAS-driven malignancies.
Typical applications include quantitative assessment of signaling dynamics via Western blot for phosphorylated ERK and AKT, phenotypic analyses of cell proliferation (MTT, BrdU) and anchorage-independent growth in soft agar, and migration/invasion assays using transwell or wound-healing formats. The cells are also suited for in vivo xenograft tumor growth studies and high-throughput drug screening to validate small-molecule inhibitors targeting the RAS pathway. For further details, please contact Ascent Research.
