E2F6 Knockout HAP1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the HAP1 cell line, designed for loss-of-function studies of the E2F6 gene. The polyclonal format provides a heterogeneous pool of cells with targeted disruption of the E2F6 locus, enabling robust functional genomics analyses without the bottlenecks of clonal selection. This population model is suitable for investigating E2F6-mediated transcriptional repression and cell cycle control in a near-haploid human background.
HAP1 is a near-haploid human cell line originally derived from KBM-7 chronic myeloid leukemia cells, exhibiting an adherent, fibroblast-like morphology. Its near-haploid karyotype simplifies genetic manipulation and reduces the complexity of gene editing outcomes, making it a favored platform for genetic screens and knockout studies. The HAP1 cell line retains intact core cellular machinery for cell cycle regulation, DNA replication, and epigenetic control, providing a physiologically relevant context for studying the functions of transcriptional regulators like E2F6.
E2F6 encodes a transcriptional repressor that, upon dimerization with DP1, binds E2F-responsive promoter elements and recruits polycomb repressive complexes, including EZH2, SUZ12, EED, RING1, RYBP, and BMI1, to catalyze histone H3 lysine 27 trimethylation (H3K27me3) and silence target gene expression. This activity negatively regulates cell cycle progression by repressing key downstream targets such as Cyclin E1, CDC6, MCM3, and MCM5, which are critical for G1/S transition and DNA replication initiation. Through these interactions, E2F6 integrates signals from the E2F transcription factor network with epigenetic silencing machinery to maintain cellular quiescence and control proliferation.
In the HAP1 background, disruption of E2F6 provides a valuable system to dissect the role of polycomb-mediated gene silencing in cell cycle regulation and oncogenic processes. The near-haploid nature of HAP1 cells ensures that knockout effects are not masked by a second allele, allowing clear interpretation of functional outcomes related to E2F6 loss. This model is particularly suited for high-throughput genetic screens and mechanistic studies that require a simplified genomic context.
Researchers can employ this polyclonal knockout population in a wide array of assays. Western blotting and RT-qPCR can confirm loss of E2F6 protein or transcript, while flow cytometry-based cell cycle analysis enables assessment of altered proliferation kinetics. ChIP-qPCR experiments can probe changes in histone modifications and transcription factor occupancy at E2F target genes, and co-immunoprecipitation studies can examine disrupted interactions with DP1, polycomb components, or other binding partners. Proliferation assays and reporter assays further facilitate functional analyses of E2F6-mediated repression. Additionally, RNA-seq can be used for transcriptome-wide assessment of derepression effects. This product is ideal for cancer research, epigenetics, and cell cycle studies. For further details, please contact Ascent Research.
