The DPCD Knockout HEK293T Polyclonal Cells represent a CRISPR/Cas9-mediated gene disruption pool targeting the DPCD locus in HEK293T cells, providing a genetically heterogeneous cell population with loss-of-function across multiple clones. This polyclonal knockout model is optimized for functional genomics and pathway dissection in a widely used human embryonic kidney epithelial host, enabling robust investigation of ciliogenesis and centrosome biology without clonal selection bias.
HEK293T cells are a well-characterized subline of the HEK293 human embryonic kidney epithelial line, stably expressing the SV40 large T-antigen to enhance episomal replication of transfected plasmids. This host background facilitates high-level recombinant protein expression and efficient virus production, making it a versatile workhorse for biochemical, cell biological, and pharmacological assays. The adherent epithelial phenotype supports studies requiring polarized cell architecture, relevant to primary cilium research.
DPCD encodes a centrosomal protein critical for centriole cohesion, basal body docking, and primary cilium assembly. Mechanistically, DPCD interacts with key centrosomal proteins such as CEP63 and CP110, and its loss disrupts recruitment of intraflagellar transport (IFT) components, including IFT88 and IFT172, to the basal body. Consequently, primary cilia fail to form, leading to dampened Hedgehog signaling characterized by reduced expression of PTCH1 and GLI1, and dysregulated cell cycle progression. Thus, DPCD is an essential node connecting centrosome integrity, ciliary trafficking, and developmental signaling cascades.
In the HEK293T epithelial model, knockout of DPCD abolishes serum starvation?Cinduced ciliogenesis, providing a tractable system to dissect molecular requirements for cilium formation and Hedgehog pathway activation. The loss of primary cilia mimics cellular phenotypes observed in ciliopathies such as primary ciliary dyskinesia, enabling detailed mechanistic studies and therapeutic target validation. Furthermore, the interaction between DPCD and the BBSome complex, via BBS4 and BBS5, highlights its relevance to studying broader ciliary trafficking defects.
Typical research applications include fluorescence microscopy?Cbased quantification of cilia frequency using antibodies against acetylated ??-tubulin and ARL13B, biochemical analysis of Hedgehog signaling components by RT-qPCR for GLI1 and PTCH1 mRNA, flow cytometric cell cycle profiling, and transcriptomic approaches like RNA-seq to map genome-wide consequences of ciliary loss. This polyclonal knockout population is well-suited for high-content screening to identify small molecules that restore ciliogenesis or modulate Hedgehog output, accelerating drug discovery efforts for ciliopathies. For further details, please contact Ascent Research.
