The HRAS Knockout HCT 116 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population, generated from HCT 116 colorectal carcinoma cells by targeted disruption of the HRAS gene. This heterogeneous pool of edited cells avoids clonal bias and provides a robust model for investigating HRAS-dependent signaling. The polyclonal format ensures comprehensive coverage of gene disruption events.
HCT 116 is a human colorectal carcinoma epithelial cell line with microsatellite instability-high (MSI-H) status and an endogenous KRAS G13D activating mutation. It harbors wild-type p53 and exhibits adherent, epithelial growth. These features make HCT 116 a widely used model for colorectal cancer research, particularly for studying oncogenic Ras signaling and drug responses.
HRAS encodes a small GTPase that cycles between inactive GDP-bound and active GTP-bound states. Receptor tyrosine kinases, such as EGFR and FGFR, activate HRAS through the SOS1/GRB2 guanine nucleotide exchange factor complex. GTP-bound HRAS engages multiple effectors: RAF1 initiates the MEK1/2?CERK1/2 cascade, phosphorylating ERK1/2 to activate ELK1 and MYC transcription factors, which upregulate cyclin D1; PI3K triggers AKT and mTOR signaling to enhance cell survival and protein synthesis; RALGDS and PLCE1 contribute to additional downstream pathways. Negative regulation is provided by the NF1 GTPase-activating protein. In the HCT 116 background, where KRAS G13D drives constitutive MAPK activity, HRAS knockout allows isolation of HRAS-specific functions and potential cross-talk with PI3K/AKT signaling.
Disruption of HRAS in the KRAS-mutant HCT 116 background enables the dissection of Ras isoform-specific functions and may reveal compensatory signaling mechanisms. This polyclonal knockout model is well-suited for investigating HRAS contributions to proliferation, survival, and drug resistance, particularly toward inhibitors targeting RAF, MEK, or PI3K/mTOR. Researchers can assess alterations in phospho-ERK and phospho-AKT levels, cell cycle distribution, and apoptosis, confidently attributing effects to gene disruption rather than clonal variation. The model supports both biochemical and functional analyses within a relevant cancer context.
These cells can be employed in a range of assays, including western blotting, RT-qPCR, proliferation and colony formation assays, flow cytometry for cell cycle and apoptosis, phospho-ERK ELISA, co-immunoprecipitation, and xenograft tumor growth studies. Applications include oncogene signaling research, drug screening, and functional genomics. For detailed product information, please contact Ascent Research.
