The DNASE1L1 Knockout SK-HEP-1 Polyclonal Cells represent a CRISPR/Cas9-mediated gene-disrupted polyclonal cell population in which the DNASE1L1 locus has been targeted to abolish gene function. This product provides a loss-of-function model for studying the role of the secreted endonuclease DNASE1L1 in liver sinusoidal endothelial biology.
The host SK-HEP-1 cell line, originally derived from the ascites of a liver adenocarcinoma patient, displays hallmark endothelial characteristics and serves as a well-established model of human liver sinusoidal endothelial cells (LSECs). These cells are responsible for the fenestrated endothelial lining of hepatic sinusoids, facilitating blood-liver exchange, endocytic clearance, and immune regulatory functions.
DNASE1L1 encodes a calcium- and magnesium-dependent secreted deoxyribonuclease that specifically degrades double-stranded DNA. Its transcription is upregulated by p53, and it acts downstream of apoptotic signals mediated by caspase-3 and BAX/cytochrome c release. In the context of neutrophil extracellular trap (NET) formation, DNASE1L1 degrades NET-derived DNA, a process that involves citrullination of histone H3 by PAD4 and the activity of neutrophil elastase. The enzyme also reduces the availability of DNA autoantigens, thereby limiting anti-DNA autoantibody production. Key upstream regulators include TNF-??, IL-6, all-trans retinoic acid, and the vitamin D receptor, placing DNASE1L1 at the intersection of apoptosis, innate immunity, and inflammatory signaling.
In the liver sinusoidal microenvironment, LSECs are continuously exposed to circulating DNA from apoptotic cells and NETs. Knockout of DNASE1L1 in SK-HEP-1 cells abrogates their ability to clear extracellular DNA, leading to accumulation of DNA debris that can trigger innate immune sensors and pro-inflammatory cytokine release, such as IL-6 and TNF-??. This model is therefore particularly relevant for investigating the contribution of impaired endothelial DNA clearance to the pathogenesis of systemic lupus erythematosus and other autoimmune disorders, as well as hepatic inflammation and fibrosis.
Researchers can employ these polyclonal knockout cells to dissect the role of endothelial DNASE1L1 in DNA degradation, NET resolution, and inflammatory signaling. Representative applications include evaluating DNA accumulation by immunofluorescence and TUNEL assays, measuring extracellular DNA degradation activity, assessing NET breakdown via co-culture experiments, and profiling cytokine responses by ELISA. Additionally, the model may be used to screen therapeutic agents that aim to restore DNA clearance or modulate downstream inflammatory pathways. For further technical details, please contact Ascent Research.
