E2F8 Knockout HEK293T Polyclonal Cells comprise a CRISPR/Cas9-edited polyclonal knockout cell population derived from the HEK293T human embryonic kidney cell line, designed for functional loss-of-function studies of the atypical E2F transcription factor E2F8. This polyclonal knockout model enables investigation of E2F8-dependent gene regulation without clonal selection, providing a heterogeneous population that reflects diverse editing outcomes while maintaining robust knockout efficiency. The polyclonal format is suitable for pooled functional genomics applications, including CRISPR-based genetic screening, and reduces the clonal variation often observed in monoclonal lines.
HEK293T cells are a widely utilized human embryonic kidney epithelial cell line stably expressing the SV40 large T-antigen, which facilitates episomal replication of plasmids containing the SV40 origin of replication and enhances heterologous protein production. Their adherent epithelial morphology, rapid growth kinetics, and high transfection efficiency make them a preferred host for lentivirus production, protein overexpression, and a broad range of cell-based assays. These characteristics ensure reliable and reproducible experimental outcomes when studying gene function in a well-characterized and readily manageable cellular context.
E2F8 encodes an atypical E2F transcription factor that functions primarily as a transcriptional repressor by forming heterodimers with DP proteins (TFDP1, TFDP2) and binding to E2F consensus sites in target gene promoters. E2F8 activity is regulated by upstream signals including MYC, E2F1, p53, and canonical Wnt signaling, and is post-transcriptionally modulated by miR-129-5p. It represses key cell cycle regulators such as CCNE1 and CCNA2, thereby controlling G1/S transition and proliferation. Additionally, E2F8 suppresses angiogenic factors including VEGFA, MMP2, and MMP9, and interacts with repressive chromatin modifiers like EZH2. Through these interactions, E2F8 integrates proliferative and angiogenic programs downstream of CDK2, RB1, and p53 pathways.
In HEK293T cells, CRISPR-mediated disruption of E2F8 derepresses its downstream targets, leading to upregulation of cyclins CCNE1 and CCNA2, which promote cell cycle progression and increased proliferation. Simultaneously, elevated VEGFA expression enhances pro-angiogenic potential. This loss-of-function model thus mimics aspects of oncogenic signaling frequently observed in hepatocellular carcinoma, breast, and lung cancers, where E2F8 dysregulation contributes to tumor growth and angiogenesis. The HEK293T background further permits high-efficiency lentiviral packaging for subsequent delivery of rescue constructs or reporter systems, enabling sophisticated mechanistic dissection of E2F8 functions.
This polyclonal knockout cell product is ideally suited for a range of research applications, including cell cycle analysis via flow cytometry, proliferation assays (MTS/MTT), apoptosis evaluation, and migration/invasion studies. The upregulation of VEGFA makes the cells a valuable tool for angiogenesis research, including tube formation assays and VEGF signaling studies. Transcriptomic changes can be assessed by RT-qPCR and RNA-seq, while protein interactions and chromatin occupancy can be examined using co-immunoprecipitation and ChIP-qPCR. The polyclonal format also supports pooled CRISPR screens for genetic modifier interrogation. For further technical details or custom inquiries, please contact Ascent Research.
