The INO80E Knockout HEK293T Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population targeting the INO80E gene in HEK293T cells. This loss-of-function model disrupts expression of INO80E, a subunit of the INO80 chromatin remodeling complex, enabling investigation of nucleosome dynamics and DNA repair. The polyclonal format provides a heterogeneous pool of edited cells, minimizing clonal bias and suitable for population-based functional assays.
HEK293T is a human embryonic kidney epithelial cell line immortalized with adenovirus 5 DNA and expressing SV40 large T antigen. This background supports high-level protein expression, efficient viral packaging, and robust transient transfection, making it a workhorse for recombinant protein production and gene delivery applications. Its epithelial origin also allows studies of kidney cell biology.
INO80E is a component of the multi?subunit INO80 ATP?dependent chromatin remodeling complex, which mobilizes nucleosomes along DNA to regulate transcription, replication, and repair. The complex is activated upstream by DNA damage signaling via ATM/ATR kinases and cell cycle regulators. INO80E directly interacts with core complex members, including INO80, ACTR5, ACTR8, RUVBL1, RUVBL2, and the transcription factor YY1, to facilitate nucleosome repositioning and histone variant H2A.Z exchange. This activity promotes access for DNA repair factors such as RAD51 and BRCA1 and influences gene expression programs. Loss of INO80E destabilizes the complex, impairing chromatin remodeling at damage sites and leading to defective repair and transcriptional dysregulation.
In the HEK293T context, INO80E knockout provides a powerful model to dissect how chromatin dynamics influence genome stability. The cell line??s ease of manipulation allows combinatorial studies with ectopic expression of INO80E variants or reporter constructs to monitor repair kinetics. This model is especially relevant for elucidating molecular mechanisms underlying cancer predisposition and neurodevelopmental disorders linked to INO80 complex dysfunction.
Research applications include chromatin remodeling studies, DNA repair pathway analysis, and epigenetic regulation. Typical assays are Western blotting for INO80E and complex subunits, ChIP for nucleosome occupancy, immunofluorescence for ??H2AX and DNA repair foci, comet assay for DNA damage, cell viability assays after genotoxic challenge, RNA-seq for transcriptional profiling, and co-immunoprecipitation to assess complex integrity. These polyclonal knockout cells thus support functional genomics and drug discovery programs targeting chromatin modifiers. For further information, contact Ascent Research.
