The C17orf100 Knockout HEK293T Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal knockout cell population with targeted disruption of the C17orf100 gene in the HEK293T human embryonic kidney cell line. This heterogeneous knockout model arises from CRISPR/Cas9-mediated gene disruption, resulting in loss of functional C17orf100 protein. The polyclonal format provides a diverse genetic background that is useful for robust phenotypic analysis without selection for a single clone.
HEK293T cells are an adenovirus 5-transformed derivative of HEK293 cells that stably expresses the SV40 large T-antigen, enabling high transfection efficiency and robust recombinant protein expression. This cell line is extensively utilized for lentiviral production and functional genomics studies. As a kidney epithelial cell line, HEK293T retains the ability to form primary cilia upon serum withdrawal, offering a suitable context for investigating cilia-related processes.
C17orf100 (HEATR9) encodes a microtubule-associated HEAT repeat protein that localizes to centrosomes and basal bodies, where it is essential for primary cilium assembly and ciliogenesis. It functions by organizing centriolar satellites and facilitating intraflagellar transport (IFT), processes critical for ciliary formation and signaling. Transcriptional regulation of C17orf100 is mediated by RFX family transcription factors, E2F1, and FOXJ1, establishing its role in ciliogenic programs. Within the centrosome, C17orf100 interacts with PCM1, CEP164, and CEP152, and is required for proper localization of downstream factors including IFT88, PCM1, CEP290, and BBS4. Disruption of C17orf100 compromises primary cilium integrity and attenuates hedgehog signaling, which depends on a functional cilium for signal transduction.
In the HEK293T background, knockout of C17orf100 permits detailed examination of centrosome biology and ciliogenesis. HEK293T cells can be induced to form primary cilia by serum starvation, enabling direct assessment of ciliary defects via immunofluorescence microscopy for ciliary markers. This model allows investigation of how centrosomal protein complexes coordinate ciliary assembly, without confounding effects from oncogenic signaling common in many cancer lines.
Applications include ciliopathy disease modeling for disorders such as Joubert syndrome and Meckel-Gruber syndrome, analysis of primary cilium biogenesis, hedgehog signaling studies, and drug screening for ciliopathies. Common assays involve serum starvation-induced ciliogenesis, immunofluorescence for acetylated tubulin and gamma-tubulin, western blotting, RT-qPCR, and co-immunoprecipitation. For detailed product information and support, please contact Ascent Research.
