The CCDC9B Knockout HEK293T Polyclonal Cells are a CRISPR/Cas9-edited population of human embryonic kidney cells with targeted disruption of the CCDC9B gene. This polyclonal format comprises a heterogeneous pool of edited cells, providing a versatile loss-of-function model without clonal selection. It enables investigation of CFAP91 function within the widely used HEK293T background, facilitating dissection of the nexin-dynein regulatory complex (N-DRC) in a human cellular context.
HEK293T cells derive from human embryonic kidney cells transformed with adenovirus 5 DNA and stably express the SV40 large T antigen, permitting episomal plasmid replication and high transfectability. They are a cornerstone for transient protein expression, viral vector production, and biochemical assays due to rapid growth and ease of handling. This well-characterized line provides a robust platform for generating knockout populations with minimal confounding factors.
CCDC9B encodes CFAP91, a coiled-coil protein integral to the N-DRC in ciliary and flagellar axonemes, where it stabilizes microtubule doublet interactions and couples dynein motor forces to axonemal bending. CFAP91 is transcriptionally regulated by the ciliogenic factor FOXJ1 and interacts with N-DRC components DRC1, DRC3, and DRC4, axonemal dyneins DNAI1 and DNAH5, and downstream radial spoke and tubulin proteins. This positions CFAP91 as a central scaffold essential for microtubule-based motility and ciliary function.
While HEK293T cells do not naturally form motile cilia, they can be induced to assemble primary cilia under appropriate conditions, allowing investigation of CFAP91’s role in axonemal stability. Their high transfectability facilitates rescue experiments and interaction studies, making the polyclonal knockout ideal for biochemical analyses such as co-immunoprecipitation and immunofluorescence. The pool’s genetic diversity reduces clonal biases, supporting robust population-level assays.
This model supports research into primary ciliary dyskinesia, Kartagener syndrome, and male infertility by enabling functional dissection of N-DRC components and screening for motility modulators. Key assays include immunofluorescence for ciliary markers (ARL13B, acetylated ??-tubulin), high-speed video microscopy for beat frequency, co-immunoprecipitation of N-DRC complexes, quantitative RT-PCR for FOXJ1 target genes, and Western blot analysis. For further details, contact Ascent Research.
