The GPATCH11 Knockout HAP1 Polyclonal Cells product provides a CRISPR/Cas9-edited polyclonal population in which the GPATCH11 gene has been disrupted in the near-haploid HAP1 cell line. This polyclonal format preserves the genetic heterogeneity inherent to pooled knockout pools, offering a robust loss-of-function model for studying GPATCH11-dependent processes without the constraints of single-cell clonal selection. The disruption is generated via CRISPR/Cas9-mediated gene targeting, producing a functional knockout suitable for a wide range of cell-based assays.
HAP1 cells are a haploid genetic model derived from the KBM-7 chronic myeloid leukemia line, characterized by a near-haploid karyotype that eliminates the complications of diploid redundancy. This unique genomic configuration enables clear genotype-phenotype correlations and facilitates genetic interaction studies, making HAP1 an ideal host for functional genomics and knockout investigations. The cells maintain key signaling pathways relevant to hematological malignancy research while providing a simplified genetic background for mechanistic dissection.
GPATCH11 encodes a G-patch domain-containing protein implicated in the DNA damage response, specifically in homologous recombination (HR) repair of double-strand breaks. Mechanistically, GPATCH11 functions downstream of the DNA damage sensors ATM and ATR, promoting RAD51 loading onto processed breaks to initiate recombinational repair. It interacts directly with key HR mediators including BRCA1, BRCA2, and PALB2, facilitating RAD51 nucleoprotein filament formation at damage sites. Disruption of GPATCH11 abrogates efficient HR, leading to persistence of unrepaired breaks and genomic instability.
In the HAP1 haploid context, GPATCH11 knockout provides a penetrant model to dissect HR pathway dependencies without interference from a second functional allele. This system is particularly valuable for examining epistatic relationships with other repair factors and for high-throughput genetic modifier screens aimed at identifying synthetic lethal interactions. The polyclonal nature of the knockout ensures that the observed phenotypes reflect the average effect of gene disruption across a diverse pool, enhancing the robustness of quantitative assays such as drug sensitivity profiling.
Researchers can employ these cells for detailed DNA repair pathway analysis, including assessment of RAD51 foci formation by immunofluorescence, ??H2AX immunoblotting to monitor double-strand break signaling, and comet assays to evaluate DNA damage accumulation. The model additionally supports clonogenic survival assays after genotoxic stress, co-immunoprecipitation studies to map GPATCH11 interaction networks, and genome-wide synthetic lethality screens in cancer drug discovery. With its broad applicability to cancer biology and genomic instability research, this product serves as a versatile tool for both mechanistic studies and translational applications. For further information and technical support, please contact Ascent Research.
