DNASE2 Knockout MES-OV Polyclonal Cells are a CRISPR/Cas9-edited polyclonal population derived from the MES-OV human ovarian carcinoma cell line, featuring targeted disruption of the DNASE2 gene. This heterogeneous knockout pool offers a robust loss-of-function model for studying the lysosomal endonuclease DNASE2 without clonal selection, preserving genetic diversity crucial for unbiased functional studies in ovarian cancer biology.
The MES-OV cell line is a widely used model of high-grade serous ovarian cancer, an aggressive epithelial malignancy. These cells exhibit dysregulated apoptosis and lysosomal activity characteristic of ovarian tumors, making them an ideal host for investigating DNASE2 in the context of cancer cell death pathways. The MES-OV background provides a relevant system for exploring how lysosomal DNA degradation impacts tumor progression and drug response.
DNASE2 is a lysosomal endonuclease responsible for degrading DNA in acidic environments, playing a critical role in apoptosis and erythrocyte enucleation. Its activity is activated by caspase-mediated apoptotic signaling and lysosomal acidification, and it interacts with the mannose-6-phosphate receptor for lysosomal targeting. DNASE2 functions downstream of caspase-3 and caspase-7, which process key substrates during cell death, and its action generates degraded DNA fragments found in TUNEL-positive nuclei and apoptotic bodies. The enzyme also collaborates with lysosomal hydrolases to complete DNA catabolism. Knockout of DNASE2 disrupts this degradation, causing accumulation of undigested DNA that can engage DNA damage sensors, potentially feeding back on upstream regulators like BAX and cytochrome c release following lysosomal membrane permeabilization.
In MES-OV ovarian cancer cells, loss of DNASE2 creates a valuable model for dissecting how impaired DNA clearance alters cell death execution and drug sensitivity. The accumulation of undigested DNA may mimic conditions seen in autoimmune disorders and affect tumor immunogenicity. This knockout system allows investigation of the interplay between lysosomal function and apoptosis, shedding light on resistance mechanisms to chemotherapy that rely on intact DNA degradation pathways.
These polyclonal knockout cells are suitable for a range of research applications, including ovarian cancer apoptosis studies, drug sensitivity testing, and dissecting lysosome-dependent cell death. Key assays include TUNEL staining and DNA fragmentation analysis to assess DNA degradation defects, Western blotting for caspase-3/7 activation, flow cytometry for Annexin V/PI staining, RT-qPCR for DNASE2 expression, and immunofluorescence for lysosomal markers. Moreover, the cells can be used to explore the role of DNASE2 in erythroid differentiation-related processes and in innate immune responses to self-DNA. For further details, please contact Ascent Research.
