DMXL1 Knockout Huh-7 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population designed for the study of DMXL1 gene function in a human hepatic background. The product consists of a heterogeneous pool of Huh-7 cells with targeted disruptions in the DMXL1 locus, enabling loss-of-function analysis without clonal selection. This polyclonal format provides a diverse genetic background for population-level functional studies.
The Huh-7 cell line is a well-characterized human hepatocellular carcinoma model with epithelial morphology, originally derived from a liver tumor of a 57-year-old Japanese male. Extensively used to investigate hepatocyte metabolism, liver cancer biology, and drug metabolism, Huh-7 cells offer a robust platform for dissecting the roles of endosomal trafficking and signaling pathways in liver tumorigenesis.
DMXL1 encodes a scaffold protein that, together with WDR7, recruits and regulates vacuolar H+-ATPase (V-ATPase) at synaptic vesicles and endosomes. This complex is critical for endolysosomal acidification, which drives neurotransmitter loading, hormone secretion, and Notch receptor activation. DMXL1-dependent acidification facilitates ??-secretase-mediated Notch1 cleavage, releasing the intracellular domain to activate transcription. DMXL1 also interacts with Rab3 GTPases and SNARE complex components, linking it to membrane fusion and vesicle trafficking. Loss of DMXL1 impairs V-ATPase regulation, disrupts endosomal pH, and attenuates Notch signaling, affecting synaptic communication and endocrine function.
In Huh-7 hepatoma cells, DMXL1 knockout allows interrogation of the interplay between endosomal trafficking, Notch signaling, and hepatocellular carcinoma. Aberrant Notch activity contributes to liver cancer stem cell maintenance and chemoresistance; DMXL1 loss may attenuate these oncogenic programs. Disrupted endolysosomal function can also affect autophagy and metabolic adaptability, processes often dysregulated in liver cancer. Thus, this model helps dissect DMXL1??s role in sustaining malignancy, including effects on drug sensitivity.
This polyclonal knockout population supports diverse assays: Western blotting and RT-qPCR confirm target disruption and pathway expression; immunofluorescence and LysoTracker staining visualize endosomal defects; flow cytometry tracks cell cycle and apoptosis; MTT assays assess viability and drug sensitivity; ???secretase assays probe Notch activation; and migration/invasion assays evaluate metastatic potential. Suitable for functional genomics and cancer drug discovery. For further details, contact Ascent Research.
