The INO80E Knockout HeLa Polyclonal Cells consist of a CRISPR/Cas9-edited polyclonal population of HeLa cells engineered for targeted disruption of the INO80E gene. This loss-of-function model abolishes INO80E function across a heterogeneous pool of edited alleles, providing a reproducible genetic background for investigating INO80E-dependent mechanisms. The polyclonal format preserves overall cellular heterogeneity while eliminating the need for clonal isolation, thereby reducing selection-associated artifacts and ensuring robust phenotype assessment in downstream applications.
The HeLa cell line, an immortalized human cervical adenocarcinoma line positive for human papillomavirus type 18 (HPV-18), serves as a widely utilized model in cancer biology and gene function studies. These cells exhibit rapid proliferation, facile genetic manipulation, and well-documented genomic and proteomic landscapes, making them particularly suitable for research into DNA damage response and chromatin biology. Their transformation by HPV-18 oncoproteins results in abrogation of p53 and pRB tumor suppressor pathways, a context that underscores the relevance of this knockout model for studying genome maintenance in the absence of normal checkpoints.
INO80E encodes a subunit of the INO80 chromatin remodeling complex, which uses ATP hydrolysis to remodel chromatin. The complex is crucial for DNA double-strand break repair via homologous recombination by evicting H2A.Z from damaged loci and enabling access for repair machinery. INO80E interacts with core subunits INO80, ARP5, ARP8, ACTR5, and the RUVBL1/RUVBL2 ATPases. Upstream kinases ATM and ATR activate the complex upon DNA damage, while transcription factors E2F1 and MYC regulate INO80E expression. Downstream, the complex facilitates BRCA1 and RAD51 loading, controls H2A.Z dynamics, and modulates p21-mediated cell cycle arrest. Thus, INO80E knockout disrupts DNA repair, transcription, and checkpoint signaling, promoting genomic instability.
In the HeLa context, loss of INO80E exacerbates genomic stress, providing a powerful system to dissect chromatin remodeling-dependent DNA repair pathways. The absence of functional p53 highlights reliance on INO80E for managing replication stress and DNA damage, potentially revealing synthetic lethal relationships exploitable in cancer therapy. This model is invaluable for studying INO80 complex contributions to repair pathway choice, chromatin dynamics, and cell survival following genotoxic insult. Moreover, the ease of large-scale culture and transfection supports high-content screening and systems-level analyses of INO80E networks.
Typical applications include quantifying DNA damage via ??H2AX and 53BP1 foci immunofluorescence, measuring repair efficiency by comet assay and RAD51 focus formation, and performing transcriptome profiling via RNA-seq to identify INO80E-regulated genes. Colony formation assays and flow cytometric cell cycle analysis reveal impacts on proliferation and checkpoint control. Drug sensitivity screens, particularly with PARP inhibitors and other DNA-damaging agents, identify therapeutic vulnerabilities. ChIP-qPCR for H2A.Z and histone modification mapping further elucidate mechanistic roles. For further details or technical support regarding this product, please contact Ascent Research.
