The ATRIP Knockout HAP1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal population carrying target-gene disruption in the HAP1 near-haploid human cell line. This heterogeneous knockout pool provides a versatile loss-of-function model for studying ATRIP-dependent signaling without clonal selection, enabling robust functional analyses in a genetically tractable haploid background.
The HAP1 cell line originates from KBM-7 chronic myeloid leukemia cells and maintains a near-haploid karyotype, which simplifies genetic studies by allowing single-allele disruptions to manifest as complete functional knockouts. Its cancer-derived origin offers a relevant context for investigating tumor-suppressor pathways, DNA damage responses, and drug sensitivity mechanisms.
ATRIP is a critical cofactor that forms a stable complex with the ATR kinase. Upon replication stress or DNA damage, RPA-coated single-stranded DNA recruits the ATRIP-ATR heterodimer, which is then activated through TOPBP1 and the 9-1-1 complex. Active ATR phosphorylates a cascade of substrates including CHK1, p53, RPA, CDC25, and histone H2AX, thereby coordinating cell cycle arrest, replication fork stabilization, and DNA repair. Additionally, ATRIP-mediated signaling regulates homologous recombination factors such as FANCD2 and SMC1, positioning ATRIP as a central node in the replication stress response and checkpoint control.
In the haploid HAP1 background, ATRIP disruption yields a clear loss of ATR-dependent signaling, making these cells an ideal platform to dissect pathway hierarchy and synthetic lethal interactions. The model is particularly suited for evaluating ATR inhibitors like VE-821 and AZD6738, and for exploring the role of ATRIP in genomic instability syndromes such as Seckel syndrome and microcephaly. Its cancer-derived lineage further enhances relevance for oncogenic stress studies and targeted therapy development.
Applications include Western blotting for p-CHK1 and ??H2AX, immunofluorescence detection of DNA damage foci, flow cytometry for cell cycle analysis, and clonogenic survival assays to measure drug sensitivity. The polyclonal knockout cells also serve as a foundational tool for high-throughput drug screens and genetic interaction studies. For expert guidance or custom assay support, contact Ascent Research.
