The DNASE2B Knockout HAP1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population targeting the DNASE2B gene in the near-haploid human HAP1 cell line. This product disrupts DNASE2B expression, creating a loss-of-function model ideal for functional studies. The polyclonal format provides a heterogeneous pool of knockout cells without the need for single-cell cloning, facilitating robust and reproducible experiments.
HAP1 cells are derived from the KBM-7 chronic myeloid leukemia line, carrying the Philadelphia chromosome, and exhibit an adherent, fibroblast-like morphology. Their near-haploid karyotype (one copy per chromosome) allows efficient genetic manipulation, as disruption of a single allele eliminates gene function, making them a powerful tool for genetic screens and functional genomics. This cell line is widely used in cancer biology, signal transduction, and drug discovery research due to its well-characterized genome and ease of culture.
DNASE2B encodes a lysosomal endonuclease responsible for DNA degradation under acidic conditions, clearing DNA from apoptotic cells and cellular debris. Loss of DNASE2B leads to accumulation of undegraded DNA, which can translocate to the cytosol and bind cGAS, activating the cGAS-STING innate immune pathway. This triggers phosphorylation of STING and TBK1, leading to IRF3 activation and transcription of type I interferons like IFN-??, along with NF-??B-mediated inflammatory responses. The pathway is regulated by upstream factors such as TFEB and TNF-??, and involves lysosomal membrane proteins LAMP1 and LAMP2 for proper enzyme localization. Thus, DNASE2B acts as a critical brake on aberrant DNA-driven immune activation.
In the HAP1 context, DNASE2B knockout offers a sensitive model for studying cytosolic DNA sensing because the haploid background reduces confounding effects from wild-type alleles. This model is particularly suited for investigating autoimmune diseases such as systemic lupus erythematosus, where defective DNA clearance contributes to pathogenesis. It also facilitates exploration of chronic inflammation and cancer interactions, as well as high-throughput screening for inhibitors or activators of STING-dependent signaling.
Researchers can employ this knockout model for a wide range of experimental applications, including mechanistic dissection of lysosomal DNA degradation, characterization of cGAS-STING pathway dynamics, and autoimmune disease modeling. Typical assays include Western blotting for phosphorylated STING and IRF3, RT-qPCR for IFNB1 and interferon-stimulated gene expression, ELISA for secreted IFN-??, and immunofluorescence for colocalization of dsDNA with lysosomal markers. Additional approaches like RNA-seq, flow cytometry, and cGAS activity assays are compatible. For more information, please contact Ascent Research.
