The DNASE2 Knockout HEK293T Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population in which the human DNASE2 gene has been disrupted, generating a loss-of-function model for studying lysosomal DNA degradation and innate immune signaling. This polyclonal pool of edited HEK293T cells offers a heterogeneous knockout background suitable for bulk functional analyses without the limitations of single-cell clonal selection, enabling researchers to capture population-level responses to DNASE2 deficiency.
The HEK293T host cell line is derived from adenovirus-5 transformed human embryonic kidney epithelial cells and stably expresses the SV40 large T antigen, which promotes episomal replication of transfected plasmids and high-level recombinant protein expression. These characteristics make HEK293T cells a widely utilized platform for transient transfection, viral packaging, and signal transduction studies, providing a reliable and versatile cellular background for introducing targeted genetic modifications.
DNASE2 encodes a lysosomal endonuclease that degrades DNA from apoptotic cells and erythroid nuclei, preventing cytoplasmic accumulation of self-DNA. Loss of DNASE2 leads to undegraded DNA engaging the cytosolic sensor cGAS, which produces cGAMP to activate STING. STING recruits TBK1, which phosphorylates IRF3 and IRF7, inducing type I interferon expression. This signaling cascade is regulated by upstream factors such as TFEB, which controls lysosomal biogenesis, and erythropoietin during erythropoiesis, while the M6P receptor mediates lysosomal targeting of DNASE2.
HEK293T cells express key components of the cGAS-STING pathway, making them suitable for studying DNA-driven innate immunity. The polyclonal DNASE2 knockout population allows examination of the immediate consequences of DNASE2 loss without clonal selection bias, preserving diverse phenotypic responses. This model is valuable for dissecting how impaired DNA degradation triggers interferon production and contributes to autoinflammatory disease signatures.
Researchers can use these cells to investigate mechanisms of autoimmune diseases such as systemic lupus erythematosus and type I interferonopathies, screen for STING pathway inhibitors, and study erythroid maturation defects. Common assays include RT-qPCR for interferon-stimulated genes, immunoblotting for phosphorylated STING and TBK1, immunofluorescence microscopy to visualize STING translocation, and DNA degradation activity measurements. For additional information, please contact Ascent Research.
