In Stock Cell Lines
Homo sapiens (Human)
Bone
Adherent
The RHOT1 Knockout U2OS Cell Line is a CRISPR/Cas9-edited human osteosarcoma model in which the RHOT1 gene is disrupted, eliminating expression of the mitochondrial Rho GTPase MIRO1. MIRO1 functions as a critical adaptor that links mitochondria to the microtubule motor proteins KIF5 and dynein via TRAK1/TRAK2, enabling calcium- and AMPK-regulated mitochondrial transport and positioning. Loss of MIRO1 impairs mitophagy initiation by preventing PINK1/Parkin-mediated degradation. This cell line supports investigation of mitochondrial dynamics, mitophagy, and metabolic reprogramming in cancer. Applications include live-cell imaging, co-immunoprecipitation, ATP assays, and drug screening targeting mitochondrial quality control relevant to neurodegeneration and oncology.
NDUFB11 Knockout SK-HEP-1 Polyclonal Cells
Cat. No. ARG15580
ATF1 Knockout Hela Polyclonal Cells
Cat. No. ARG20936
IDI2 Knockout HAP1 Polyclonal Cells
Cat. No. ARG22684
IL6ST Knockout A549 Polyclonal Cells
Cat. No. ARG34118
EVPL Knockout MES-OV Polyclonal Cells
Cat. No. ARG6032
MRC-5
Cat. No. ARP1002
The RHOT1 Knockout U2OS Cell Line is a CRISPR/Cas9-edited human osteosarcoma model with targeted disruption of the RHOT1 gene, eliminating the mitochondrial Rho GTPase MIRO1. This stable knockout line enables precise loss-of-function analysis of MIRO1-dependent mitochondrial trafficking, dynamics, and mitophagy in a cancer-relevant context.
U2OS is an adherent epithelial cell line derived from a moderately differentiated human osteosarcoma, retaining osteoblastic characteristics and a stable karyotype. Its robust oxidative metabolism and well-defined responses to stress and apoptosis make it an ideal host for mitochondrial research. U2OS cells are readily transfectable and support reproducible gene-editing experiments.
RHOT1 encodes MIRO1, a mitochondrial outer membrane GTPase that couples mitochondria to microtubule motors via TRAK1/TRAK2 adaptor proteins, enabling bidirectional transport by KIF5 and cytoplasmic dynein. MIRO1 activity is calcium-regulated through EF-hand domains and modulated by AMPK phosphorylation, linking transport to cellular energy status. Upon mitochondrial damage, PINK1/Parkin phosphorylate and ubiquitinate MIRO1, targeting it for degradation to halt transport and initiate mitophagy. Downstream effects of MIRO1 loss include impaired mitochondrial distribution, reduced local ATP supply, disturbed calcium handling, and enhanced cytochrome c release during apoptosis. The MIRO1 network intersects with mitochondrial fusion (MFN1/MFN2) and fission (DRP1) pathways, coordinating transport with organelle morphology.
In U2OS cells, RHOT1 ablation causes perinuclear mitochondrial clustering, diminishing peripheral trafficking essential for cell migration and invasion. This redistribution may heighten apoptotic sensitivity and shift metabolic flux, providing a system to examine oxygen consumption versus glycolysis. The knockout line serves as a clean backdrop for studying mitophagy defects via the PINK1/Parkin pathway, particularly under hypoxia or nutrient stress. Given MIRO1??s links to cancer progression and Parkinson??s disease, the model is applicable to both oncology and neurodegeneration research.
The cell line supports live-cell imaging of mitochondrial movement, co-immunoprecipitation of MIRO1 motor complexes, and metabolic flux analysis. Assays for ATP levels, mitochondrial membrane potential, and apoptosis enable functional profiling of RHOT1 deficiency. Western blotting and RT-qPCR confirm knockout and assess changes in DRP1, OPA1, and VDAC1. It is also suited for high-content drug screens targeting mitochondrial dynamics or mitophagy. For technical inquiries, please contact Ascent Research.
