The CCDC91 Knockout HEK293T Polyclonal Cells comprise a CRISPR/Cas9-mediated gene-disrupted polyclonal population designed to ablate CCDC91 function. As a heterogeneous pool of knockout cells, this product avoids clonal artifacts and is ideal for population-averaged biochemical and cell-based assays requiring consistent loss-of-function across a large cell cohort.
HEK293T is a human embryonic kidney cell line immortalized with adenovirus 5 E1A/E1B and stably expressing SV40 large T antigen. These cells are widely adopted for high-level recombinant protein production and lentiviral/retroviral packaging due to their high transfection efficiency. Though not typically ciliated in standard culture, HEK293T’s robust signaling machinery and ease of genetic manipulation make it a practical host for dissecting ciliary transport and signaling pathways when combined with forced ciliation or as a surrogate system.
CCDC91 functions as a scaffold adaptor essential for intraflagellar transport (IFT) and primary cilium biogenesis. It directly binds core IFT-B subunits IFT20, IFT88, and IFT52, as well as BBSome components BBS4 and BBS5, to orchestrate the trafficking of ciliary membrane proteins including Smoothened (SMO) and ARL13B. CCDC91 expression is driven by ciliogenic transcription factors RFX3 and FOXJ1, and its activity is downstream of Hedgehog (Hh) pathway activation, where SHH binding to Patched1 (PTCH1) derepresses SMO, triggering a signaling cascade that culminates in GLI1/2/3 transcription factor activation. Disruption of CCDC91 uncouples IFT from ciliary signal transduction, leading to impaired Hh and Wnt pathway outputs.
In the HEK293T context, CCDC91 knockout provides an accessible model to study the molecular pathology of ciliopathies such as Joubert syndrome, Meckel syndrome, and nephronophthisis. The polyclonal knockout pool enables researchers to examine how heterogeneous loss of CCDC91 impacts IFT particle integrity, ciliary membrane composition, and the transcriptional activity of GLI proteins without necessitating complex ciliogenesis induction protocols. This makes it a versatile platform for genetic and pharmacological intervention studies.
Representative applications include high-content immunofluorescence screening for ciliary markers (acetylated tubulin, ARL13B), co-immunoprecipitation to assess IFT complex formation, dual-luciferase reporter assays for Hedgehog pathway activity, RT-qPCR quantification of GLI target gene expression, and western blot analysis of IFT/BBSome protein levels. Additionally, cell migration assays can evaluate downstream functional effects of ciliary signaling loss. These polyclonal knockout cells are suited for ciliogenesis mechanism investigation, ciliary trafficking modulator screening, drug discovery for polycystic kidney disease, and hedgehog inhibitor profiling. For technical inquiries or ordering, please contact Ascent Research.
