The CCDC171 Knockout HEK293T Polyclonal Cells are a CRISPR/Cas9-edited polyclonal cell population featuring targeted disruption of the human CCDC171 gene across a heterogeneous pool of HEK293T cells. This loss-of-function model is designed for studying CCDC171 in a widely used host background. The polyclonal format avoids single-clone selection, providing a sampling of diverse editing events suitable for population-level functional assays and high-throughput screening applications.
HEK293T cells, derived from human embryonic kidney, express the SV40 large T antigen, enabling episomal plasmid replication and facilitating high-efficiency transient transfection and recombinant protein production. Under serum-starvation conditions, these cells can form primary cilia, making them a pertinent model for ciliogenesis research. Their robust proliferation and compatibility with imaging and biochemical workflows underscore their utility as a chassis for investigating ciliary assembly pathways.
CCDC171 encodes a predicted coiled-coil domain scaffold protein participating in cilium assembly and microtubule-based processes. It functions downstream of ciliogenic transcription factors such as RFX proteins and FOXJ1, and interacts with intraflagellar transport components including IFT88 and IFT140, along with BBSome elements. These interactions suggest CCDC171 coordinates cargo loading onto IFT machinery, thereby influencing axonemal construction and ciliary signaling. Disruption may impair cilium formation or downstream signal transduction.
In HEK293T cells, CCDC171 knockout provides a system to directly interrogate its necessity for ciliogenesis. Because ciliation can be induced, the model permits temporal dissection of assembly steps and signaling defects. The polyclonal cell pool captures a range of functional impairment, mimicking tissue heterogeneity. Beyond ciliary roles, the model may reveal contributions of CCDC171 to cell cycle control or cytoskeletal organization, given the multifunctionality of ciliary proteins.
Typical applications include immunofluorescence-based quantification of ciliary markers (ARL13B, acetylated tubulin), co-immunoprecipitation for protein interaction mapping with IFT88 or IFT140, and RT-qPCR profiling of ciliary gene expression. High-content screening for cilia modulators can leverage the polyclonal pool??s diversity. These cells also support phospho-signaling analysis via western blotting and flow cytometric cell cycle studies, aiding ciliopathy research and functional genomics. For further details, contact Ascent Research.
