INO80C Knockout HAP1 Polyclonal Cells constitute a CRISPR/Cas9-edited polyclonal knockout cell population designed for constitutive disruption of the INO80C gene in the human near-haploid HAP1 cell line. This product provides a heterogeneous pool of gene-edited cells, enabling functional interrogation of INO80C without the need for single-cell cloning. The polyclonal format preserves biological variability while establishing a loss-of-function model, making it well-suited for pooled genetic screens and population-based functional assays.
HAP1 cells are derived from the KBM-7 chronic myeloid leukemia (CML) cell line, which was isolated from a patient in blast crisis. HAP1 exhibits a near-haploid karyotype, retaining a single copy of most chromosomes except for a disomic region of chromosome 15, which simplifies genetic knockout studies by eliminating confounding effects from a second functional allele. Despite its haploid nature, HAP1 maintains functional DNA repair, cell cycle control, and chromatin remodeling pathways, offering a streamlined genetic background to study genes involved in genome maintenance and leukemia biology.
INO80C is a core subunit of the INO80 ATP-dependent chromatin remodeling complex, which slides and evicts nucleosomes at gene promoters and DNA damage sites. Within the complex, INO80C interacts with INO80, RUVBL1, RUVBL2, ACTR5 (ARP5), IES2, and the histone variant H2A.Z to coordinate chromatin dynamics. The complex is recruited to double-strand breaks by ??H2AX, a modification catalyzed by ATM and ATR kinases, where it facilitates homology-directed repair by promoting nucleosome clearance and enabling the recruitment of RAD51 and BRCA1. INO80C also contributes to transcriptional regulation by modulating nucleosome positioning at target gene promoters, influenced by upstream signals from the cell cycle machinery and DNA damage checkpoints.
Disruption of INO80C in the HAP1 near-haploid context is expected to compromise INO80 complex activity, resulting in impaired DNA repair, altered gene expression programs, and increased sensitivity to genotoxic stress. This model is particularly valuable for dissecting the contributions of chromatin remodeling to CML and blast crisis progression, as well as to broader mechanisms of genomic instability in cancer. The combination of INO80C loss and the near-haploid background allows for clean phenotypic assessment of DNA damage signaling and repair pathway dependencies.
This polyclonal knockout cell population enables a diverse set of research applications, including genetic interaction screens to identify synthetic lethal partners, functional dissection of DNA damage response using ??H2AX foci formation, comet assays, and homology-directed repair reporter assays, and chromatin analyses via ChIP-qPCR to assess nucleosome occupancy. Transcriptional consequences can be explored by RNA-seq, while cell cycle effects are analyzed by flow cytometry. The cells are also applicable to drug sensitivity profiling and synthetic lethality screens in the context of CML. For additional details or purchasing information, please contact Ascent Research.
