The DNASE2 Knockout K-562 Polyclonal Cells product consists of a polyclonal population of K-562 cells that have undergone CRISPR/Cas9-mediated disruption of the DNASE2 gene, resulting in a loss-of-function knockout model. This polyclonal knockout cell population is generated from the human K-562 chronic myeloid leukemia cell line, providing a heterogeneous pool of edited cells suitable for pooled functional studies. The CRISPR/Cas9 editing introduces targeted gene disruption across the cell population, allowing researchers to study the collective consequences of DNASE2 deficiency without relying on a single clone. As a knockout model, these cells enable investigation of DNASE2-dependent biological processes in a hematopoietic context.
The host K-562 cell line is a human hematopoietic model derived from the pleural effusion of a CML patient in blast crisis, harboring the BCR-ABL oncogene. K-562 cells are widely utilized for studies of cell proliferation, differentiation, and apoptosis due to their erythroleukemic properties. Their suspension growth and responsiveness to differentiation-inducing agents facilitate high-throughput assays.
DNASE2 encodes a lysosomal acid deoxyribonuclease that degrades DNA from apoptotic cells and erythroblast nuclei within phagolysosomes, preventing immunogenic DNA accumulation. Its expression is regulated by TFEB and MITF in response to phagocytic and apoptotic stimuli, and it is targeted to lysosomes via the mannose-6-phosphate receptor. DNASE2 collaborates with other lysosomal hydrolases like cathepsins for efficient DNA clearance. In the absence of DNASE2, undegraded DNA escapes lysosomes, activating cGAS, which stimulates STING-dependent TBK1/IRF3 phosphorylation and IFN-?? production. This pathway links DNASE2 to suppression of innate immune responses and erythroid enucleation.
In K-562 cells, DNASE2 knockout creates a model to investigate DNA-induced innate immune signaling and its impact on cell fate. These cells are primed for erythroid differentiation and apoptosis, allowing dissection of how DNASE2 deficiency promotes cGAS-STING activation, interferon responses, and defective nuclear clearance. The polyclonal population recapitulates heterogeneous responses to DNA damage, autophagy modulation, and inflammatory gene expression, making it ideal for studying autoinflammatory and erythropoietic disorders.
Key applications include monitoring cGAS-STING pathway activation via phospho-STING/IRF3, IFN-?? ELISA, and RNA-seq of interferon-stimulated genes. The cells support erythroid differentiation assays with flow cytometry, lysosomal immunofluorescence, and DNA degradation measurements. Apoptosis kinetics and autophagy flux can also be evaluated. This model enables research into autoimmune pathology, DNA damage responses, and phagosomal clearance mechanisms in a hematopoietic context. For inquiries, contact Ascent Research.
