The DNAL1 Knockout Huh-7 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population designed to interrogate DNAL1 function in hepatocellular carcinoma. These cells harbor targeted gene disruptions at the DNAL1 locus, circumventing clonal artifacts and providing a heterogeneous model for studying ciliary biology. It enables robust assessment of DNAL1 loss in processes such as ciliary beat generation and Hedgehog pathway modulation. This product is ideal for exploring axonemal dynein mechanics and cilia-dependent signaling in a liver context.
The Huh-7 cell line was established from a well-differentiated hepatocellular carcinoma of a 57-year-old Japanese male in 1982. It maintains key hepatic characteristics and, under serum starvation, can assemble primary cilia, making it a valuable system for studying ciliary function in liver cancer. The DNAL1 knockout in this tumorigenic background thus provides a clinically relevant model for examining how ciliary defects intersect with oncogenic pathways.
DNAL1 encodes a light chain of the axonemal outer dynein arm, a multimeric motor complex that includes DNAH5, DNAI1, DNALI1, CCDC114, and ARMC4. Its expression is transcriptionally regulated by the master ciliogenic factors FOXJ1, RFX2, and RFX3. DNAL1 is essential for ATP-dependent microtubule sliding, generating the force for ciliary beat and fluid flow. This mechanical activity is a prerequisite for proper Hedgehog signal transduction. Interacting directly with other arm subunits, DNAL1 contributes to dynein structural integrity; its disruption therefore compromises ciliary motility, cell migration, and downstream signaling, placing it at a critical node in ciliopathy research.
In Huh-7 cells, DNAL1 knockout enables dissection of ciliary motility’s role in liver tumor biology. Despite the frequent loss of cilia in cancer, Huh-7 retains ciliogenic capacity, allowing direct assessment of how outer dynein arm defects affect hepatocellular carcinoma behavior. This model is suited for studying cilia-dependent Hedgehog signaling, cell migration, and tumor microenvironmental interactions. Furthermore, it can be employed to screen for modulators of ciliary function with therapeutic potential in liver cancer.
Common experimental uses include immunofluorescence staining for ciliary markers such as acetylated ??-tubulin and ARL13B to assess ciliation, high-speed video microscopy to quantify ciliary beat frequency, and biochemical assays like western blotting for DNAL1 and RT-qPCR for ciliogenesis-related transcripts. Functional endpoints can be evaluated through wound healing assays to measure cell migration. This model is particularly valuable for primary ciliary dyskinesia research and for dissecting cilia-dependent pathways in hepatocellular carcinoma. For additional technical information, please contact Ascent Research.
