The RHOA Knockout LN-229 Cell Line is a precisely engineered CRISPR/Cas9-edited knockout cell line derived from the human LN-229 glioblastoma cell line. This product features targeted disruption of the RHOA gene, which encodes the Ras homolog family member A (RhoA) small GTPase, a central regulator of actin cytoskeleton dynamics. The knockout model enables loss-of-function studies of RhoA-dependent signaling pathways. It is supplied as a validated cell line suitable for oncological, neurological, and pharmacological research applications requiring native tumor cell context.
LN-229 is a well-characterized cell line established from a patient with glioblastoma multiforme (GBM), the most common and aggressive primary brain tumor. These cells retain key features of GBM, including invasive capacity, tumorigenicity in xenograft models, and aberrant activation of mitogenic and survival pathways. LN-229 has been extensively used as an in vitro and in vivo model system to investigate GBM biology, therapeutic resistance, and tumor?Cmicroenvironment interactions. The RHOA knockout thus offers a genetically defined derivative for dissecting RhoA??s contribution to malignant phenotypes in a clinically relevant GBM background.
RhoA functions as a molecular switch, cycling between inactive GDP-bound and active GTP-bound states. Guanine nucleotide exchange factors (GEFs) like VAV, LARG, and DBL promote GTP loading downstream of diverse receptors including GPCRs, RTKs such as EGFR, integrins, and cytokine receptors. Upon activation, RhoA binds effectors including Rho-associated coiled-coil kinase (ROCK) and mammalian diaphanous (mDia). ROCK phosphorylates LIM kinase (LIMK) and myosin light chain (MLC), resulting in cofilin inhibition and actomyosin contraction, respectively, which drive stress fiber formation and focal adhesion maturation. mDia stimulates actin polymerization through profilin, while RhoA also engages protein kinase N (PKN) to activate NF-??B and the MRTF/SRF transcriptional complex to control cytoskeletal gene expression. RhoGDI and GTPase-activating proteins such as p190RhoGAP provide negative regulation. This network integrates extracellular signals to govern cell shape, adhesion, migration, and proliferation.
In glioblastoma, RhoA is frequently overactivated due to oncogenic receptor signaling and mutations, driving tumor invasion, proliferation, and therapy resistance. The LN-229 RHOA knockout cell line enables dissection of ROCK-dependent and -independent mechanisms underlying cytoskeletal remodeling and 3D invasion. It serves as an ideal model for evaluating therapeutic inhibitors of the RhoA?CROCK axis, such as fasudil or Y-27632, and for exploring crosstalk with PI3K/AKT and Wnt signaling.
Representative assays include Western blotting for phospho-MLC and total RhoA, G-LISA activation assays, Transwell migration and invasion studies, and immunofluorescence staining for F-actin and vinculin. The model supports RNA-seq analysis to define RhoA-dependent transcriptomes and drug sensitivity screening with ROCK inhibitors such as fasudil. Co-immunoprecipitation of RhoA with ROCK or mDia validates disrupted interactions. The RHOA Knockout LN-229 Cell Line is an essential tool for GBM and Rho signaling research and preclinical drug discovery. For more information, please contact Ascent Research.
