The E2F5 Knockout HEK293T Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population with targeted disruption of the human E2F5 gene in the HEK293T host cell line. This heterogeneous pool enables loss-of-function studies of the E2F5 transcriptional repressor without clonal isolation, making it suitable for pooled functional genomics and high-throughput screening applications.
HEK293T cells are a derivative of human embryonic kidney epithelium that stably express the SV40 large T antigen. This modification enhances episomal plasmid replication, leading to amplified protein expression and increased viral production. These cells are widely used for transient and stable protein production, virus packaging, and as a versatile platform for molecular and cell biology research.
E2F5 is a member of the E2F transcription factor family that predominantly functions as a transcriptional repressor. It forms complexes with pocket proteins p130 and p107, and dimerizes with DP partners (TFDP1, TFDP2) to bind E2F response elements. This repressor complex recruits histone deacetylases HDAC1/2 and SWI/SNF chromatin remodeling factors to silence cell cycle genes such as CCNA2, CCNE1, CDC6, and MCM family members. Upstream, E2F5 activity is regulated by cyclin-dependent kinases CDK4/6 and CDK2 through phosphorylation of pRB family proteins, integrating signals from TGF-beta and MYC pathways. Additionally, E2F5 influences apoptosis regulators like BIRC5, positioning it as a key node controlling cell cycle exit and differentiation.
In the HEK293T background, disruption of E2F5 is expected to relieve repression of its target genes, potentially altering cell cycle dynamics and proliferation. Given the host cell??s reliance on robust transcriptional machinery for high-level expression and viral production, the knockout model allows dissection of the repressive arm of the RB/E2F pathway in a widely used, easily transfectable epithelial system. This provides insights into how E2F5 balances quiescence and proliferation.
This polyclonal knockout pool is suited for cell cycle research, cancer biology, and drug target validation. Applications include Western blot and RT-qPCR to confirm E2F5 depletion and target gene deregulation, RNA-seq for transcriptomic profiling, and flow cytometry for cell cycle analysis. Functional assays such as MTT, BrdU, Annexin V, and migration assays assess phenotypic consequences, while co-immunoprecipitation and ChIP-qPCR probe E2F5 interactions and promoter occupancy. The cells support mechanistic studies, pooled screens, and gene therapy research. For more information, contact Ascent Research.
