The DNASE2 Knockout Huh-7 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the human Huh-7 hepatocellular carcinoma line. This product provides a robust loss-of-function model for DNASE2, a lysosomal endonuclease critical for DNA degradation under acidic conditions. The polyclonal format ensures a heterogeneous mixture of cells with targeted gene disruptions, facilitating broad functional analyses.
Huh-7 is a well-differentiated hepatocellular carcinoma cell line established from a liver tumor in a 57-year-old Japanese male. These liver epithelial cells retain active metabolic and protein synthesis capabilities, making them a valuable model for hepatic biology, cancer research, and drug metabolism studies. Their tumorigenic origin and epithelial characteristics provide a relevant context for examining pathways that connect lysosomal function, innate immunity, and oncogenesis.
DNASE2 encodes a lysosomal endonuclease that cleaves DNA in acidic environments, essential for degrading DNA from apoptotic cells and cellular debris. The enzyme is regulated upstream by TFEB and mTORC1, which coordinate lysosomal biogenesis, and it interacts with cathepsins and lysosomal membrane proteins such as LAMP1 and LAMP2. Loss of DNASE2 activity results in accumulation of undegraded DNA, which can activate endosomal TLR9 or the cytosolic cGAS-STING pathway, triggering production of inflammatory cytokines. This molecular axis underscores DNASE2??s role in preventing DNA-driven autoimmunity and chronic inflammation.
In Huh-7 cells, DNASE2 knockout disrupts lysosomal DNA degradation, creating a potent model to study innate immune activation in a hepatic context. Given the liver??s role in clearing apoptotic debris and exposure to microbial DNA, accumulation of undegraded DNA can chronically engage STING-dependent interferon responses, mirroring aspects of liver inflammation seen in autoimmune disorders. Additionally, the interplay between lysosomal stress and mTORC1 signaling in these metabolically active cells offers insights into metabolic-immune crosstalk relevant to hepatocellular carcinoma and other liver diseases.
This knockout model supports diverse research applications, including dissecting DNA sensing pathways, investigating apoptosis and phagocytosis mechanisms, and screening compounds that inhibit cGAS-STING or TLR9 signaling. Key experimental approaches include immunofluorescence to monitor DNA accumulation, TUNEL and flow cytometry for apoptosis assessment, ELISA for cytokine secretion profiling, and phagocytosis assays to evaluate clearance of apoptotic cells. For additional details or customization requests, please contact Ascent Research.
