The ANKS6 Knockout HEK293T Polyclonal Cells consist of a CRISPR/Cas9-edited polyclonal population with targeted gene disruption at the ANKS6 locus. As polyclonal knockout cells, this product offers a genetically diverse loss-of-function model without the biases associated with single-cell cloning, allowing assessment of gene-dosage effects and population-level phenotypic variation. Derived from Homo sapiens, these cells provide a versatile platform for studying the ciliary protein ANKS6 in an immortalized cell background.
HEK293T cells are human embryonic kidney epithelial cells immortalized by adenoviral E1A and E1B gene expression and further engineered to harbor the SV40 large T antigen. This modification supports high-copy episomal replication of transfected plasmids, enabling efficient protein overexpression, lentivirus production, and transient gene silencing or expression studies. The adherent epithelial morphology and rapid doubling time of HEK293T facilitate routine culture and high-throughput screening, while their renal origin maintains baseline expression of many kidney-related genes, providing contextual relevance for ciliopathy research.
ANKS6 (Ankyrin Repeat and SAM Domain-Containing Protein 6) is an essential component of the ciliary transition zone, where it assembles into a supramolecular complex with NPHP3, INVS, and NEK8 to negatively regulate kidney cystogenesis. This complex functions downstream of ciliary signaling inputs, including those from primary cilia deflection and Wnt ligand stimulation, and is transcriptionally regulated by RFX factors. Loss of ANKS6 disrupts ciliary homeostasis, leading to aberrant activation of both canonical Wnt/??-catenin and Hippo/YAP-TAZ pathways, with ??-catenin stabilization and YAP/TAZ nuclear accumulation driving deregulated epithelial proliferation and planar cell polarity defects. Interactors such as ANKS3 and intraflagellar transport proteins further underscore its nodal role in ciliary signal integration.
Within HEK293T cells, ANKS6 knockout creates a tractable system for investigating ciliary protein function and downstream signaling cascades. Although HEK293T normally grow as monolayers, they can be adapted to three-dimensional culture conditions that promote partial polarization and lumen formation, serving as a surrogate for kidney tubular morphogenesis. The polyclonal knockout pool is particularly suited for examining population heterogeneity in Wnt and Hippo pathway responses, as variable residual ANKS6 expression across the population may mimic heterozygous states encountered in human nephronophthisis.
This knockout model supports diverse applications such as western blotting and RT-qPCR for quantifying Wnt/Hippo target gene expression, immunofluorescence for ciliary markers, and co-immunoprecipitation to probe altered protein interaction networks. Furthermore, 3D cyst growth assays and cell proliferation analyses enable functional evaluation of cystogenic potential and candidate drug screening for cystic kidney diseases including nephronophthisis type 16. For further technical details, please contact Ascent Research.
