The INO80D Knockout HAP1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population targeting the INO80D gene in the human near-haploid HAP1 cell line. This loss-of-function model enables systematic investigation of INO80D function in chromatin remodeling and DNA repair without diploid genetic complexity. The polyclonal format provides a heterogeneous pool of knockout alleles, facilitating robust phenotypic assessment while limiting clonal artifacts.
HAP1 cells derive from the KBM-7 chronic myeloid leukemia line and possess a near-haploid karyotype ideal for genetic knockout studies. Their haploid genome simplifies gene disruption, as targeting a single allele abolishes function, reducing off-target effects and accelerating knockout population generation. Extensively characterized, HAP1 cells are widely adopted in functional genomics, drug screening, and mechanistic studies. Combined with CRISPR/Cas9-mediated INO80D ablation, they provide a genetically clean, experimentally tractable platform for dissecting chromatin remodeling and DNA repair pathways.
INO80D encodes a core subunit of the INO80 chromatin remodeling complex, which uses ATP to slide nucleosomes and exchange histones, regulating DNA accessibility. The complex is recruited to DNA damage sites by upstream kinases ATM and ATR, with its activity also influenced by the YY1 transcription factor. INO80D interacts with INO80, ACTL6A/B, RUVBL1/2, and ARP5 to maintain complex integrity. Downstream, the complex deposits H2A.Z, modulates histone modifications, and controls genes such as HOX clusters and cell cycle regulators. INO80D disruption thus impairs nucleosome dynamics, hinders DNA double-strand break repair (reducing RAD51 loading and causing persistent ??-H2AX foci), and causes transcriptional dysregulation, compromising genome stability.
In the HAP1 near-haploid background, INO80D knockout yields a definitive loss-of-function phenotype, enabling clear dissection of its role in the DNA damage response within a leukemic context. This model is valuable for studying how chromatin remodelers contribute to cancer cell survival and therapy resistance. INO80D absence sensitizes cells to genotoxic stress, making these cells suitable for synthetic lethality drug screens with chromatin remodeling inhibitors. Epigenetic consequences can be linked to oncogenic gene expression changes.
These polyclonal cells support a range of assays, including Western blotting, ChIP-qPCR for histone modifications at HOX loci, and immunofluorescence for ??-H2AX foci. Transcriptomics via RNA-seq identifies dysregulated genes, while comet and cell viability assays measure DNA damage responses. Applications span chromatin biology, DNA repair, cancer epigenetics, and drug target validation. For additional information, please contact Ascent Research.
