The DRICH1 Knockout HEK293T Polyclonal Cells constitute a CRISPR/Cas9-edited polyclonal knockout cell population derived from the widely used HEK293T host cell line. This product provides a heterogeneous mixture of cells carrying disruptions in the DRICH1 gene, which encodes a core component of the nexin-dynein regulatory complex (N-DRC) essential for ciliary motility. The polyclonal format allows researchers to study loss-of-function effects without relying on a single clonal isolate, offering a more robust representation of the knockout phenotype across a population of gene-edited cells.
HEK293T cells are an adherent, epithelial-like derivative of the human embryonic kidney HEK293 line, stably expressing the SV40 large T antigen, which facilitates episomal plasmid replication and high-level transient protein expression. Upon serum starvation, these cells can assemble primary cilia, making them a tractable model for studying ciliogenesis and ciliary protein function. This host background supports a wide array of biochemical and cell biological assays, including immunofluorescence imaging of ciliary markers, protein?Cprotein interaction studies, and functional analyses of ciliary components.
DRICH1 (CCDC164) is a coiled-coil domain protein that serves as a core subunit of the N-DRC, a large complex that links peripheral microtubule doublets in the axonemal shaft and coordinates dynein-driven microtubule sliding. DRICH1 interacts with other DRC subunits such as DRC1, DRC2, DRC3, and DRC7, as well as with dynein heavy chains including DNAH5 and DNAH11. This protein complex functions downstream of transcription factors like RFX family members and FOXJ1, which promote motile ciliogenesis, and is upregulated in response to serum starvation-induced ciliary assembly. Loss of DRICH1 disrupts N-DRC integrity, leading to impaired regulation of dynein motor activity and defective ciliary beat coordination, a hallmark of motile ciliopathies such as primary ciliary dyskinesia (PCD).
In the HEK293T background, which supports formation of primary cilia upon serum withdrawal, the DRICH1 knockout population provides a physiologically relevant system to dissect N-DRC composition and function. Although HEK293T cells typically produce non-motile primary cilia, overexpression of key motile ciliogenesis factors or utilization of specialized culture conditions can induce axonemal motility components, enabling studies of DRICH1??s role in dynein regulation. This model thus bridges the gap between simple biochemical analysis and more complex ciliary functional studies, allowing investigation of N-DRC assembly, stability, and interaction with microtubule structures.
Researchers can employ this knockout product for a variety of applications, including co-immunoprecipitation assays to map N-DRC subunit interactions, western blotting to monitor complex stability, and immunofluorescence microscopy using ciliary markers (acetylated tubulin, ARL13B) to assess ciliogenesis and ciliary length. The polyclonal nature also supports pooled functional screens and cell migration assays that may be influenced by ciliary signaling. Moreover, electron microscopy can be utilized to examine axonemal ultrastructure changes in the absence of DRICH1. For more information on custom gene-edited cell products, please contact Ascent Research.
