The DMXL1 Knockout 786-O Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population with targeted disruption of the DMXL1 gene in the 786-O human renal cell carcinoma line. This product provides a heterogeneous pool of edited cells that avoids clonal artifacts, enabling functional studies of DMXL1 in a population context. The polyclonal format is generated by CRISPR/Cas9-mediated gene disruption, avoiding single-cell cloning and making it suitable for assays requiring robust biological replicates.
The 786-O cell line is derived from a primary clear cell renal adenocarcinoma and carries a VHL mutation leading to constitutive HIF1A stabilization. This mimics the VHL loss prevalent in ccRCC, establishing an epithelial model for investigating hypoxia-driven oncogenic pathways. The line’s genetic background makes it valuable for studying genes involved in renal cancer progression.
DMXL1 encodes Rabconnectin-3??, a scaffold protein that coordinates Notch receptor activation and V-ATPase assembly. It interacts with RAB3A, RABGEF1, and RABGAP1L, and forms complexes with V-ATPase subunits (ATP6V0A1, ATP6V1A) to regulate lysosomal acidification. In Notch signaling, DMXL1 facilitates transmission downstream of ligands DLL1, DLL4, JAG1, JAG2, interacting with NOTCH1, and promoting NICD/RBPJ/MAML1-mediated transcription of HES1, HEY1, and MYC. DMXL1-dependent lysosomal function also influences mTORC1 activity, linking intracellular trafficking to growth signals. Knockout disrupts these processes, impairing Notch target gene expression and lysosomal acidification.
In VHL-mutant 786-O cells, DMXL1 knockout provides a model to dissect crosstalk between Notch, mTORC1, and HIF1A pathways in ccRCC. Constitutive HIF1A activation may intersect with DMXL1-mediated acidification and trafficking, affecting proliferation and survival. Loss of DMXL1 can attenuate Notch targets and mTORC1 signaling, potentially revealing dependencies in VHL-deficient renal cancer cells. This cell system enables investigation of how DMXL1-dependent pathways contribute to oncogenic phenotypes under hypoxic mimicking conditions.
Applications include mechanistic studies of Notch signaling, V-ATPase function, and Rab GTPase-mediated trafficking. Assays such as Western blotting for NICD, HES1, and mTOR markers; RT-qPCR for Notch targets; immunofluorescence for lysosomal pH; flow cytometry; and co-immunoprecipitation of DMXL1 partners (NOTCH1, RAB3A) are enabled. Migration, invasion, and drug sensitivity assays can assess treatment responses. The polyclonal nature supports population-level screening and functional genomics. For further details, please contact Ascent Research.
