The DYNLT1 Knockout Huh-7 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the human Huh-7 hepatocellular carcinoma line. This product provides a loss-of-function model for DYNLT1, which encodes the dynein light chain Tctex-1 subunit essential for cytoplasmic dynein motor function. The polyclonal format provides a heterogeneous mixture of edited cells, suitable for population-level assays without clonal expansion bottlenecks. CRISPR/Cas9-mediated gene disruption abrogates DYNLT1 protein expression, enabling dissection of dynein-dependent pathways in liver cancer contexts.
Huh-7 is a well-differentiated epithelial hepatocellular carcinoma cell line established from a Japanese male liver tumor. It is extensively used for hepatocellular carcinoma research, drug metabolism and toxicology studies, and as a permissive host for hepatitis C virus replication. The cell line retains hepatocyte-specific functions and signaling networks, making it a relevant model for liver cancer biology and viral hepatitis. Its adherent growth and high transfection efficiency support diverse genetic manipulation and functional assays.
DYNLT1 encodes Tctex-1, a light chain of cytoplasmic dynein that mediates minus-end-directed transport along microtubules. It interacts with dynein intermediate chains (DYNC1I1/2), heavy chain (DYNC1H1), and the dynactin complex (DCTN1), and is regulated by assembly factors including Lis1 (PAFAH1B1), NudE/NudEL (NDEL1), and cell cycle kinases such as CDK1. DYNLT1 is critical for mitotic spindle organization, TGF-beta receptor endosomal trafficking, and apoptosis. In the TGF-beta pathway, dynein-dependent trafficking of TGFBR1 influences SMAD2/3 phosphorylation, while interactions with BCL-2 family members tie DYNLT1 to cell survival. Thus, DYNLT1 connects cytoskeletal dynamics, growth factor signaling, and programmed cell death.
In Huh-7 cells, DYNLT1 knockout disrupts cytoplasmic dynein function, impairing minus-end-directed transport of organelles and protein complexes. This leads to mitotic defects, altered TGF-beta receptor trafficking, and attenuated SMAD2/3 signaling, which may suppress epithelial-mesenchymal transition (EMT) and invasion. Additionally, mitochondrial positioning and BCL-2-regulated apoptosis can be perturbed, potentially altering drug sensitivity. These effects are particularly relevant in hepatocellular carcinoma, where dynein-dependent processes contribute to tumor progression, chemoresistance, and viral replication.
These polyclonal DYNLT1 knockout cells enable investigation of dynein-dependent transport in liver cancer, including host factors supporting HCV and HBV replication. They are suitable for dissecting TGF-beta-driven EMT through biochemical, imaging, and migration/invasion assays, as well as for screening dynein inhibitors as potential therapeutics. Representative assays include Western blotting for dynein components and SMADs, immunofluorescence for mitotic spindles and cargo localization, cell cycle analysis by flow cytometry, apoptosis assays, co-immunoprecipitation to assess dynein complex integrity, and RNA-seq. The model also supports drug sensitivity testing and studies of mitochondrial dynamics. For further details or to discuss your research requirements, please contact Ascent Research.
