CCDC181 Knockout hTERT-RPE1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the hTERT-RPE1 human retinal pigment epithelial cell line, engineered for targeted disruption of the CCDC181 gene. This loss-of-function model provides a mixed genetic background that reflects population-level effects of CCDC181 ablation, avoiding clonal artifacts and enabling robust study of gene function in a non-tumorigenic, ciliated epithelial context. The polyclonal nature captures heterogeneous editing outcomes, ideal for examining overall phenotypic consequences and signaling perturbations without the selection bias of single-cell clones.
The host hTERT-RPE1 cell line is a telomerase-immortalized, non-transformed retinal pigment epithelial model that retains key epithelial polarity, a functional primary cilium, and critical RPE functions such as phagocytosis of photoreceptor outer segments, nutrient transport, and maintenance of the blood-retinal barrier. Its well-characterized ciliogenesis program and stable ciliary architecture make it a gold-standard system for investigating centrosomal and ciliary proteins, including those linked to Hedgehog signaling and intraflagellar transport. This background ensures physiological relevance for studying retinal biology and ciliopathy-associated mechanisms.
CCDC181 encodes a coiled-coil domain protein that localizes to centrosomes and primary cilia, where it is implicated in ciliogenesis and ciliary signaling. Transcription of CCDC181 is regulated by FOXJ1 and RFX transcription factors, key drivers of ciliary gene programs. The protein interacts with centrosomal and ciliary components including CEP290, PCM1, and tubulin, positioning it within the centrosome-cilium interface. Disruption of CCDC181 is expected to impair ciliary assembly or stability, leading to defective processing of GLI3 into its repressor form and reduced expression of Hedgehog target genes GLI1 and PTCH1, thereby attenuating pathway output. Additional pathway components affected may include SMO, ARL13B, and acetylated tubulin, all critical for cilium-dependent signal transduction.
In the hTERT-RPE1 context, loss of CCDC181 likely perturbs primary cilium length, ciliation frequency, and Hedgehog pathway responsiveness, mirroring defects seen in ciliopathies and retinal degenerative diseases. RPE cells rely on proper ciliary function for cellular homeostasis and intercellular signaling, and disruption of centrosomal-ciliary proteins can compromise photoreceptor support and visual cycle integrity. This polyclonal knockout model thus recapitulates disease-relevant cellular phenotypes and enables dissection of CCDC181??s role in centrosome biology and cilium-mediated signaling within a well-defined, physiologically relevant retinal epithelium.
Researchers can employ this model for a wide range of applications, including cilia biology investigations, ciliopathy disease modeling, and Hedgehog signaling analysis. Representative assays include immunofluorescence microscopy for ciliary markers such as acetylated tubulin and ARL13B to assess cilia morphology and length, RT-qPCR to quantify GLI1 and PTCH1 transcript changes, and Western blotting to monitor GLI3 processing and pathway activity. Additional functional studies may utilize ciliogenesis assays under serum starvation, scratch wound migration assays, or cell cycle analysis to evaluate downstream phenotypic consequences in RPE cells. This product is also suitable for drug screening campaigns targeting ciliopathy-related pathways. For additional information or product inquiries, please contact Ascent Research.
