The DNASE2 Knockout 786-O Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the 786-O human renal cell carcinoma line, with targeted disruption of the DNASE2 gene. This loss-of-function model is designed for investigating the role of lysosomal acid deoxyribonuclease in DNA degradation and associated processes. The polyclonal format provides a heterogeneous population suited to studies where clonal variability is not desired.
The 786-O cell line is a well-established model of clear cell renal cell carcinoma (ccRCC), characterized by biallelic inactivation of the VHL tumor suppressor gene. VHL loss stabilizes hypoxia-inducible factors (HIFs), inducing a pseudo-hypoxic state that drives oncogenesis. This background makes 786-O cells relevant for studying metabolic reprogramming, angiogenesis, and stress responses in renal cancer. VHL mutation also influences lysosomal activity and autophagy, providing a context to examine links between DNA clearance and tumor cell biology.
DNASE2 encodes a lysosomal enzyme that hydrolyzes DNA under acidic conditions, essential for clearing DNA from apoptotic cells and for definitive erythropoiesis. Its activity is regulated by transcription factors TFEB and MITF, master regulators of lysosomal biogenesis, and responds to inflammatory signals. Within lysosomes, DNASE2 interacts with LAMP1 and LAMP2, collaborating with cathepsins and other hydrolases to degrade biomolecules. Catalytic action generates DNA fragments and nucleotides, linking to metabolic and immune pathways. Disruption of DNASE2 impairs lysosomal DNA degradation, potentially activating innate immune sensors like cGAS-STING.
In VHL-mutant 786-O cells, DNASE2 knockout dissects the interplay between lysosomal DNA clearance and tumor cell fitness. Loss of function causes accumulation of undigested DNA, which may trigger cGAS-STING-mediated interferon responses, affect apoptotic clearance, and alter the tumor microenvironment. Given the role of autophagy and lysosomal pathways in ccRCC, this model allows study of how impaired DNA catabolism drives inflammation, immune evasion, and therapy resistance. The polyclonal nature reflects a range of editing events, enabling analysis of gene-dose effects.
Researchers can use this model to analyze lysosomal DNA degradation kinetics, apoptosis and clearance assays, autophagy flux, and cytokine profiling for innate immune activation. Representative techniques include western blotting and RT-qPCR for DNASE2 expression, immunofluorescence for lysosomal DNA accumulation, and functional DNA degradation assays. Applications include studying cancer immune evasion, lysosomal storage-like phenotypes, and self-DNA sensing in tumors. For technical inquiries or customization, please contact Ascent Research.
