The DNAAF2 Knockout Huh-7 Polyclonal Cells product consists of a polyclonal population of Huh-7 human hepatocellular carcinoma cells engineered via CRISPR/Cas9-mediated gene disruption to introduce a loss-of-function modification in the DNAAF2 gene. This targeted gene knockout model enables the study of DNAAF2-dependent molecular mechanisms in a hepatic cellular environment without relying on single-cell clones, offering a heterogeneous yet genetically disrupted cell pool suitable for population-level functional assays.
Huh-7 is an epithelial cell line derived from a liver tumor of a 57-year-old Japanese male. This well-established model is widely utilized in hepatitis C virus replication studies and hepatic drug metabolism research due to its robust growth and relevance to liver cancer biology. The parental Huh-7 line retains key hepatocyte features and can be induced to undergo ciliogenesis under serum-deprived conditions, making it a suitable host for investigating genes involved in ciliary assembly and function.
DNAAF2 encodes a cytoplasmic co-chaperone that is essential for the preassembly of axonemal dynein arms, the molecular motors driving ciliary motility. DNAAF2 functions in concert with interacting factors such as DNAAF1, DNAAF3, and DNAAF4, and directly complexes with dynein heavy and intermediate chains to facilitate correct assembly of outer and inner dynein arms. Upstream, DNAAF2 expression is regulated by transcription factors FOXJ1, RFX2, and RFX3, which are master regulators of ciliogenesis. Downstream, DNAAF2 activity is required for dynein arm assembly and subsequent ciliary beat frequency, and its loss leads to immotile cilia. Within the broader pathway, DNAAF2 acts alongside DNAAF1, DNAAF3, dynein heavy chains, and axonemal docking complexes to ensure proper axonemal architecture and mucociliary clearance.
In the Huh-7 hepatocellular carcinoma background, this DNAAF2 knockout offers a unique tool to dissect ciliary assembly pathways in a liver-derived cell line. Although cilia are not typically prominent in hepatocytes, Huh-7 cells can be induced to form primary cilia, and disruptions in ciliary genes have been increasingly linked to cancer-related signaling, including proliferation and migration. This model therefore allows investigation of how DNAAF2 loss impacts cilia-dependent processes such as mucociliary clearance dysfunction and dynein arm assembly, as well as broader cellular phenotypes in a cancer-relevant context. It provides a relevant platform for primary ciliary dyskinesia modeling without relying on specialized respiratory cell lines.
This DNAAF2 knockout polyclonal cell population supports serum starvation-induced ciliogenesis assays with immunofluorescence detection of acetylated ??-tubulin, western blot analysis of dynein subunits, and RT-qPCR profiling of cilia-related genes. Functional assays including proliferation, migration, and cisplatin sensitivity testing extend its utility to cancer biology. Genetic interaction studies with partners such as DNAAF1 and DNAAF3 can delineate the dynein arm assembly pathway. For further technical details, please contact Ascent Research.
