The HES7 Knockout HEK293T Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal knockout cell population designed for functional studies of the HES7 gene. This product comprises a heterogeneous pool of HEK293T cells that have undergone targeted disruption of the HES7 locus via CRISPR/Cas9, resulting in a loss-of-function model suitable for investigating HES7-dependent biological processes. As a bulk polyclonal knockout, it captures a diverse set of editing outcomes, enabling robust assessment of HES7 function in a population context without single-cell clonal selection.
The host cell line, HEK293T, is an immortalized human embryonic kidney epithelial cell line that is widely employed in molecular and cellular biology due to its high transfection efficiency and reliable growth characteristics. Originally derived from HEK293 cells with stable expression of SV40 large T antigen, HEK293T offers a facile system for exogenous DNA delivery and protein expression, making it a favorable platform for studying gene regulatory networks and signaling pathways in a human cellular background.
HES7 encodes a basic helix-loop-helix (bHLH) transcriptional repressor central to the vertebrate segmentation clock, an oscillatory gene network patterning somites. Notch1 receptor activation by DLL1 ligand leads to Notch intracellular domain release, which forms a complex with RBPJ and MAML to drive HES7 transcription. HES7 then represses its own expression and downstream targets like LFNG and HES1 via delayed negative feedback, generating ultradian oscillations. Repression is partly mediated by TLE/Groucho corepressors, and oscillatory dynamics are modulated by Wnt and FGF signaling cross-talk, ensuring precise somite boundary formation.
Although HES7 is not natively expressed in HEK293T cells, this knockout provides a clean background for reconstituting clock networks via ectopic expression. HEK293T cells are adaptable for synthetic oscillators and dynamic studies, and HES7 disruption eliminates endogenous interference in reporter or inducible systems. As HES7 loss is linked to congenital vertebral defects like spondylocostal dysostosis and congenital scoliosis, this model aids in examining disease-associated variants.
Applications include RT-qPCR for oscillatory gene expression quantification, luciferase reporter assays for Notch/HES7 activity, and live-cell imaging of fluorescent oscillatory reporters. Complementary RNA-seq and western blotting can reveal transcriptomic and proteomic changes. The polyclonal nature minimizes clonal artifacts, offering a robust loss-of-function model. For technical details or ordering, contact Ascent Research.
