The IPP Knockout HAP1 Polyclonal Cells are a CRISPR/Cas9-mediated gene disruption pool targeting the IPP locus in a near-haploid human cell background. This heterogeneous polyclonal population serves as a versatile loss-of-function model for investigating IPP’s dual roles in actin cytoskeletal dynamics and DNA damage repair, without clonal expansion artifacts.
HAP1 cells are a human near-haploid chronic myeloid leukemia line derived from KBM-7, possessing a haploid karyotype except for a disomic chromosome 8 fragment and expressing the BCR-ABL1 fusion kinase. The near-haploid genome simplifies gene disruption, as a single targeted allele yields a functional null phenotype, making these cells ideal for genetic screens, drug target validation, and mechanistic studies in a hematopoietic context relevant to leukemia and cancer biology.
IPP encodes an actin-binding protein that connects cytoskeletal regulation to genomic stability. It directly interacts with actin filaments, RAD51, and Ku70 (XRCC6), thereby coordinating homologous recombination and non-homologous end joining repair pathways. Upstream regulators include RhoA, Rac1, TP53, STAT3, and NF-??B. Downstream effects involve actin polymerization, RAD51 foci formation, cell migration, and apoptosis. IPP also functions as an adaptor (KLHL27) for the Cullin3 E3 ubiquitin ligase complex, interacting with kelch-like proteins to mediate ubiquitination. These interactions integrate signals from the actin cytoskeleton and DNA damage response, influencing cell motility, DNA repair, and survival.
In the HAP1 near-haploid background, IPP disruption creates a clean genetic model for dissecting its dual functions. Loss of IPP impairs actin filament dynamics and cell motility while reducing DNA repair capacity, increasing genomic instability and modulating apoptotic thresholds. This model is especially useful for studying the interplay between migration and DNA damage responses in hematopoietic cells, with implications for understanding chemoresistance in leukemia and other cancers. The polyclonal pool enables direct functional analyses without clonal variation biases.
Applications include Western blotting for IPP, actin, RAD51, and ??H2AX; immunofluorescence for actin stress fibers and DNA damage foci; comet assays; Boyden chamber migration/invasion assays; and MTT viability screens. The cells are valuable for assessing chemosensitivity modifiers and performing co-immunoprecipitation and phospho-signaling analyses to elucidate IPP interaction networks. For further information, please contact Ascent Research.
