The DNASE2 Knockout HAP1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population, generated by disrupting the DNASE2 gene in the HAP1 human near-haploid cell line. This product provides a loss-of-function model for studying DNASE2, a lysosomal acid endonuclease critical for degrading DNA during phagocytosis and apoptosis. The polyclonal knockout format ensures representation of diverse genetic disruptions, suitable for functional genomics and screening applications without the need for single-cell cloning. The knockout cells are derived from the HAP1 parental line and exhibit fibroblast-like adherent morphology, making them amenable to standard cell culture and downstream assays.
HAP1 cells are derived from the KBM-7 chronic myeloid leukemia cell line and retain a near-haploid karyotype for most chromosomes, providing a simplified genetic background that minimizes confounding effects of diploid heterozygosity. This haploid state enables robust genotype?Cphenotype correlations in CRISPR knockout screens and functional studies. The adherent, fibroblast-like morphology facilitates high-content imaging, live-cell microscopy, and immunofluorescence-based assays. Widely used in functional genomics, HAP1 cells support efficient CRISPR/Cas9-mediated gene disruption and high transfection rates, allowing rapid generation of knockout pools for large-scale or targeted genetic perturbation studies.
DNASE2 encodes a lysosomal endonuclease acting downstream of phagocytic receptors such as TREM2 and MerTK to cleave internalized DNA into oligonucleotides under acidic conditions. This prevents cytosolic self-DNA recognition. Disrupted DNASE2 activity results in DNA accumulation, activating the cGAS-STING pathway. cGAS-derived cGAMP binds STING, triggering TBK1-dependent phosphorylation of IRF3 and type I interferon responses. Upstream regulators include transcription factors TFEB, PU.1 (SPI1), C/EBP??, and interferon-??. The nuclease is targeted to lysosomes via mannose-6-phosphate receptor and AP-1, interacting with lysosomal hydrolases. Downstream, DNASE2 activity suppresses innate immune activation, modulates TLR9 signaling, and produces salvageable deoxynucleotides. Deficiency therefore induces cGAS-STING-dependent ISG expression and autoinflammatory signaling.
In HAP1 cells, DNASE2 knockout recapitulates key aspects of lysosomal DNA degradation deficiency observed in immune and phagocytic cells. The near-haploid background accentuates phenotypes linked to innate immune signaling, as the absence of a second allele avoids compensatory regulatory effects. This makes the polyclonal knockout population particularly valuable for dissecting cGAS-STING activation dynamics and interferon response networks without clonal bias. Furthermore, HAP1 cells express the necessary machinery for lysosomal trafficking and phagocytosis-like uptake, enabling studies of DNA substrate processing and downstream innate immune responses in a simplified genetic system. The knockout can be used to explore the interplay between autophagy and lysosomal DNA clearance, as well as the cross-regulation of endolysosomal pathways.
Researchers can employ this knockout pool in high-throughput screens to identify modulators of cGAS-STING signaling, or in focused assays such as RT-qPCR for interferon-stimulated genes (e.g., IFIT1, ISG15), immunofluorescence-based visualization of lysosomal DNA accumulation, and flow cytometry for apoptotic cell clearance defects. The cells are also suitable for Western blotting to assess phospho-STING, TBK1, and IRF3, as well as RNA-seq profiling of innate immune gene expression. Additionally, they support drug screening efforts targeting autoinflammatory diseases like Aicardi-Gouti??res syndrome and systemic lupus erythematosus, where aberrant DNA sensing drives pathology. For further information or to discuss custom applications, please contact Ascent Research.
